The protocol was carried out in accordance with the principles of the Helsinki Declaration and all the methods described here have been approved by the ethics committee of Daping Hospital and written consent was obtained from the patients.
1. Instruments for operation
<ol>
	<li>Conduct all the procedures under an operating microscope with 5-10x magnification to achieve precise anastomoses and anatomic reconstruction.</li>
</ol>
2. Inclusion and exclusion criteria
<ol>
	<li>Use the following inclusion criteria: patients who experience penile glans amputation during circumcision; the injury of penile glans mainly contains glans and/or distal urethra.</li>
	<li>Use the following exclusion criteria: penile amputation; injured not during circumcision or by disposable stapler; secondary stage repair.</li>
</ol>
3. Preparation for operation
<ol>
	<li>Tie a rubber band at the root of the penis to stop bleeding in advance.</li>
	<li>Keep the amputated glans in the sterile ice-cold saline during the transfer and before replantation.
	<ol>
		<li>Once the glans is amputated, do not discard it.</li>
		<li>Immerse the amputated glans in about 10-20 mL of sterile normal saline (preferably sterile ice-cold saline), and wrap it with two layers of sterile gloves.</li>
		<li>Place it in an ice bucket or vacuum cup with ice and transfer it to a hospital where microsurgery can be performed as soon as possible.</li>
	</ol>
	</li>
	<li>Rinse and disinfect the amputated glans with iodophor three times before replantation.</li>
	<li>Give antibiotics intravenously 30 mins before surgery. Use cefuroxime sodium 0.50-0.75 g with 100 mL of 0.9% sodium chloride solution according to patient&#39;s weight.</li>
</ol>
4. Procedure
<ol>
	<li>Repairment of the amputated penile glans and frenulum if applicable.
	<ol>
		<li>Remove the amputated glans from the preserved ice bag and disinfect. Observe the morphology and integrity of the glans under the microscope. If there is fragmentation, repair it first (Figure 1A).</li>
		<li>Apply 8-0 non-absorbable thread to suture the inner cavernous incision of the glans (Figure 1B). Meanwhile, use 6-0 absorbable thread to suture the tears on the surface of the glans (Figure 1C).</li>
		<li>Trim the alongside frenulum if applicable.</li>
	</ol>
	</li>
	<li>End-to-end urethral anastomosis
	<ol>
		<li>Put the patient in a supine position.</li>
		<li>Observe and evaluate the damage of the stump of glans under the microscope (Figure 1D), and align the amputated glans and frenulum with the stump of glans. Insert an 8 Fr. Foley catheter via the urethral orifice for the drainage of urine (Figure 1E).</li>
		<li>Perform end-to-end urethral anastomosis using 6-0 absorbable thread through the intermittent suture&#160;method with precise positioning. Usually, suture one stitch at 12:00 and 6:00 o&#39;clock of the urethra, respectively at first, and then on the base of two positioning points; suture about four stitches tightly on the left and right sides, respectively in a counterclockwise or clockwise direction (Figure 1F).</li>
	</ol>
	</li>
	<li>Anastomosis of amputated glans surface and reconstruction of the frenulum
	<ol>
		<li>Suture the edge of amputated glans to the edge of the penile stump using 6-0 absorbable thread. Suture a few fixed points at 6, 12, 9, and 3 o&#39;clock at first, and then suture the space between the fixed points with a space of 1 mm for each to achieve a precise anastomosis (Figure 1G).</li>
		<li>Trim the excess inner plate of prepuce on the ventral side of the penile glans if applicable.</li>
		<li>Reconstruct the frenulum by intermittent suture to achieve a satisfactory shape if applicable (Figure 1H,I).</li>
	</ol>
	</li>
</ol>
5. Postoperative care
<ol>
	<li>Evaluate the recovery of local blood circulation by closely observing the color of the replanted penile glans. 
	NOTE: A dark red glans is expected (cherry pulp-like) and it should be observed dynamically. There are some blackish crusts on the surface after days and&#160;make sure not to remove them. Blackish crust does not mean necrosis of the amputated glans.</li>
	<li>Maintain postoperative intravenous antibiotic treatment for 2-3 days and change to oral treatment for 5-7 days.</li>
	<li>Keep the drainage of indwelling catheter unobstructed and remove it 1-2 weeks after the operation.</li>
</ol>

<ol>
	<li>Pippi Salle, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glans+amputation+during+routine+neonatal+circumcision:+mechanism+of+injury+and+strategy+for+prevention.">Glans amputation during routine neonatal circumcision: mechanism of injury and strategy for prevention.</a> Journal of Pediatric Urology. 9, 763-768 (2013).</li><li>Rizvi, S. A., Naqvi, S. A., Hussain, M., Hasan, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Religious+circumcision:+a+Muslim+view.">Religious circumcision: a Muslim view.</a> BJU International. 83, 13-16 (1999).</li><li>Weiss, H. A., Larke, N., Halperin, D., Schenker, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+of+circumcision+in+male+neonates,+infants+and+children:+a+systematic+review.">Complications of circumcision in male neonates, infants and children: a systematic review.</a> BMC Urology. 10, 2 (2010).</li><li>Harrison, N. W., Eshleman, J. L., Ngugi, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethical+issues+in+the+developing+world.">Ethical issues in the developing world.</a> BJU International. 76, 93-96 (1995).</li><li>Ceylan, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Severe+complications+of+circumcision:+an+analysis+of+48+cases.">Severe complications of circumcision: an analysis of 48 cases.</a> Journal of Pediatric Urology. 3 (1), 32-35 (2007).</li><li>Okeke, L. I., Asinobi, A. A., Ikuerowo, O. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+complications+of+male+circumcision+in+Ibadan,+Nigeria.">Epidemiology of complications of male circumcision in Ibadan, Nigeria.</a> BMC Urology. 6, 21 (2006).</li><li>Shabanzadeh, D. M., Clausen, S., Maigaard, K., Fode, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Male+circumcision+complications+-+A+systematic+review,+meta-analysis+and+meta-regression.">Male circumcision complications - A systematic review, meta-analysis and meta-regression.</a> Urology. 152, 25-34 (2021).</li><li>Aboutaleb, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reconstruction+of+an+amputated+glans+penis+with+a+buccal+mucosal+graft:+case+report+of+a+novel+technique.">Reconstruction of an amputated glans penis with a buccal mucosal graft: case report of a novel technique.</a> Korean Journal of Urology. 55 (12), 841-843 (2014).</li><li>Co&#349;kunfirat, O. K., Sayilkan, S., Velidedeoglu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glans+and+penile+skin+amputation+as+a+complication+of+circumcision.">Glans and penile skin amputation as a complication of circumcision.</a> Annals of Plastic Surgery. 43 (4), 457 (1999).</li><li>El-Bahnasawy, M. S., El-Sherbiny, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paediatric+penile+trauma.">Paediatric penile trauma.</a> BJU International. 90 (1), 92-96 (2002).</li><li>Petrella, F., Amar, S., El-Sherbiny, M., Capolicchio, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Total+glans+amputation+after+neonatal+circumcision.">Total glans amputation after neonatal circumcision.</a> Urology Case Reports. 37, 101624 (2021).</li><li>Raisin, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glans+injury+during+ritual+circumcision.">Glans injury during ritual circumcision.</a> Journal of Pediatric Urology. 16 (4), 471 (2020).</li><li>Nasr, R., Traboulsi, S. L., Abou Ghaida, R. R., Bakhach, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iatrogenic+penile+glans+amputation:+major+novel+reconstructive+procedure.">Iatrogenic penile glans amputation: major novel reconstructive procedure.</a> Case Reports in Urology. 2013, 741980 (2013).</li><li>Yosra, K., Wiem, H., Mourad, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saving+an+amputated+glans:+Role+of+winter+shunt.">Saving an amputated glans: Role of winter shunt.</a> Journal of Pediatric Urology. 16 (2), 238-240 (2020).</li><li>Manentsa, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+of+high+volume+circumcision:+glans+amputation+in+adolescents;+a+case+report.">Complications of high volume circumcision: glans amputation in adolescents; a case report.</a> BMC Urology. 19 (1), 65 (2019).</li><li>Wang, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+Replantation+of+Complete+Penile+Amputation+With+Video+Demonstration.">Microscopic Replantation of Complete Penile Amputation With Video Demonstration.</a> Urology. , (2022).</li></ol>

The authors have nothing to disclose.

For religious or therapeutic purposes, about one-quarter of men around the world have had circumcision1. The incidence of complications is far from negligible, approximately 1% to 20%3,4,5,6,7. Circumcision-related glans amputation is extremely rare and there are no exact statistics depicting its incidence. Owing to its rarity, previous...

Around 25% of men worldwide are circumcised1,2. Circumcision in China is mostly performed in childhood. Over the past decades, the improvements in surgical techniques and equipment have made circumcision less complicated, faster, and with fewer post-circumcision complications. However, the popularity of these devices has also brought new challenges.
The incidence of post-circumcision complications is around 1%-20%, mostly mild3,4,5,6,7. In a recent meta-analysis, which included 351 studies with 4,042,988 participants, the overall complication risk was 3.84% (95% confidence interval 3.35-4.37)7. Circumcision-related glans amputation is a quite rare yet devastating injury during the surgery. Such complication is conventionally treated by simple hemostasis to achieve self-healing, early gross replantation, or delayed plastic surgery8,9. However, these can result in permanent damage to penile appearance and function, along with psychologic problems, if not handled properly8,9. The prevention and treatment of glans amputation have recently developed into a challenging problem for circumcision due to the increasing public health awareness and use of various suture devices in China. There are currently no existing protocols or guidelines for the treatment of such injury, which may be due to its rarity. As a result, there is no unified understanding of the injury mechanism of glans amputations, and there is a lack of early treatment management or prevention strategies.
In this study, we reported a case series of amputated penile glans caused by dispoable stapler during&#160;circumcision, which were successfully treated by microsurgery. The&#160;technical details of microsurgery were presented by video, and their possible injury mechanisms and prevention strategies were also discussed. This protocol is applicable for early microscopic replantation of all patients with penile glans amputation caused by accidental injury.

<table><tbody><tr><td>Catheter&amp;nbsp;</td><td>Guangzhou Weili Co., Ltd</td><td>12 Fr</td><td></td></tr><tr><td>Cefuroxime sodium</td><td>Yiyi Saite, Co., Ltd</td><td>0.75 g</td><td></td></tr><tr><td>Cis-atrecu besylate</td><td>Jiangsu Dongying CO.,&amp;nbsp; Ltd</td><td>10 mg</td><td></td></tr><tr><td>Operating microscope system</td><td>Carl Zeiss Co., Ltd</td><td>OPMI VARIO 700&amp;nbsp;</td><td></td></tr><tr><td>Pentylpheptyl ether hydrochloride</td><td>Chengdu Lisi Co., Ltd</td><td>1.0 mg</td><td></td></tr><tr><td>Prolene</td><td>Ethicon, LLC</td><td>W2777/2780</td><td></td></tr><tr><td>Sufentanni citrate</td><td>Renfu Phermaceutical</td><td>50 &amp;micro;g</td><td></td></tr><tr><td>Vicryl</td><td>Ethicon, LLC</td><td>W9981</td><td></td></tr></tbody></table>

microscopic replantation of penile glans amputation due to circumcision

Circumcision using a disposable stapler is becoming quite popular in China. However, improper surgical procedures also bring the risk of penile glans amputation, which is a very rare iatrogenic genital injury. Such complication is conventionally treated by simple hemostasis to achieve self-healing, early gross replantation, or delayed plastic surgery. However, these may lead to obvious unfavorable outcomes such as amputated glans loss, necrosis, malformation healing, or urethral orifice stenosis. In the present study, we adopted microscopic replantation as an emergency approach to achieve the precise anastomoses and anatomic reconstruction of penile glans. The goal of this protocol is to present a detailed emergency management strategy with meticulous surgical skills for the penile glans amputation. The postoperative results showed that the original shape of the glans was perfectly restored with satisfactory cosmetic appearance. The micturition function was completely restored to normal without any obvious complications. There was also no significant reduction in the sensation of amputated glans area. Hence, early meticulous microscopic replantation as soon as possible is an ideal emergency management strategy for the penile glans amputation due to circumcision.

Three 7-8 years old children, whose distal penile glans were completely (2 cases) or incompletely (1 case) amputated during circumcision using a disposable stapler, were admitted to the hospital within 2-3 h after injury from June 2019 to July 2021. Physical examination confirmed that about 1/3 of the distal end of the penile glans with 3-4 mm of distal urethra was completely or incompletely amputated. In one severe case, the inner plate of prepuce and frenulum on the ventral side of the glans were also seriously damaged...

Watch this Scientific Journal Video about Microscopic Replantation of Penile Glans Amputation Due to Circumcision at JoVE.com

Microscopic Replantation of Penile Glans Amputation Due to Circumcision

The present protocol describes the emergency management of microscopic replantation of penile glans amputation due to circumcision.

Circumcision using a disposable stapler is becoming quite popular in China. However, improper surgical procedures also bring the risk of penile glans ...

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Research

JoVE Journal

Medicine

18.2K Views.  Army Medical University. The present protocol describes the emergency management of microscopic replantation of penile glans amputation due to circumcision.

Video: Microscopic Replantation of Penile Glans Amputation Due to Circumcision

Surgical Management of Meatal Stenosis with Meatoplasty

Meatal stenosis is a common urologic complication after circumcision. Children present to their primary care physicians with complaints of deviated urinary stream, difficult-to-aim, painful urination, and urinary frequency. Clinical exam reveals a pinpoint meatus and if the child is asked to urinate, he will usually have an upward, thin, occasionally forceful urinary stream with incomplete bladder emptying. The mainstay of management is meatoplasty (reconstruction of the distal urethra /meatus). This educational video will demonstrate how this is performed.

Meatoplasty, surgical management of meatal stenosis.

Meatal stenosis is a common urologic complication after circumcision.  Children present to their primary care physicians with complaints of deviated ...

A Bioluminescent and Fluorescent Orthotopic Syngeneic Murine Model of Androgen-dependent and Castration-resistant Prostate Cancer

Orthotopic tumor modeling is a valuable tool for pre-clinical prostate cancer research, as it has multiple advantages over both subcutaneous and transgenic genetically engineered mouse models. Unlike subcutaneous tumors, orthotopic tumors contain more clinically accurate vasculature, tumor microenvironment, and responses to multiple therapies. In contrast to genetically engineered mouse models, orthotopic models can be performed with lower cost and in less time, involve the use of highly complex and heterogeneous mouse or human cancer cell lines, rather that single genetic alterations, and these cell lines can be genetically modified, such as to express imaging agents. Here, we present a protocol to surgically injecting a luciferase- and mCherry-expressing murine prostate cancer cell line into the anterior prostate lobe of mice. These mice developed orthotopic tumors that were non-invasively monitored in vivo and further analyzed for tumor volume, weight, mouse survival, and immune infiltration. Further, orthotopic tumor-bearing mice were surgically castrated, leading to immediate tumor regression and subsequent recurrence, representing castration-resistant prostate cancer. Although technical skill is required to carry out this procedure, this syngeneic orthotopic model of both androgen-dependent and castration-resistant prostate cancer is of great use to all investigators in the field.

The goal of this protocol is to demonstrate the intra-prostatic injection of prostate cancer cells, with subsequent castration. Orthotopic pre-clinical models of androgen-dependent and castration-resistant prostate cancer are critical to study the disease in the context of a clinically relevant tumor microenvironment and an immunocompetent host.

Orthotopic tumor modeling is a valuable tool for pre-clinical prostate cancer research, as it has multiple advantages over both subcutaneous and ...

Cancer Research

Renal Capsule Xenografting and Subcutaneous Pellet Implantation for the Evaluation of Prostate Carcinogenesis and Benign Prostatic Hyperplasia

New therapies for two common prostate diseases, prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend critically on experiments evaluating their hormonal regulation. Sex steroid hormones (notably androgens and estrogens) are important in PrCa and BPH; we probe their respective roles in inducing prostate growth and carcinogenesis in mice with experiments using compressed hormone pellets. Hormone and/or drug pellets are easily manufactured with a pellet press, and surgically implanted into the subcutaneous tissue of the male mouse host. We also describe a protocol for the evaluation of hormonal carcinogenesis by combining subcutaneous hormone pellet implantation with xenografting of prostate cell recombinants under the renal capsule of immunocompromised mice. Moreover, subcutaneous hormone pellet implantation, in combination with renal capsule xenografting of BPH tissue, is useful to better understand hormonal regulation of benign prostate growth, and to test new therapies targeting sex steroid hormone pathways.

We describe the manufacture of compressed hormone pellets, as well as subcutaneous surgical implantation into mice. This strategy can be combined with the growth of cell and tissue xenografts under the renal capsule of athymic nude mice to evaluate hormonal carcinogenesis and regulation of benign prostate growth.

New therapies for two common prostate diseases,  prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend  critically on experiments ...

Minced Tissue in Compressed Collagen: A Cell-containing Biotransplant for Single-staged Reconstructive Repair

Conventional techniques for cell expansion and transplantation of autologous cells for tissue engineering purposes can take place in specially equipped human cell culture facilities. These methods include isolation of cells in single cell suspension and several laborious and time-consuming events before transplantation back to the patient. Previous studies suggest that the body itself could be used as a bioreactor for cell expansion and regeneration of tissue in order to minimize ex vivo manipulations of tissues and cells before transplanting to the patient. The aim of this study was to demonstrate a method for tissue harvesting, isolation of continuous epithelium, mincing of the epithelium into small pieces and incorporating them into a three-layered biomaterial. The three-layered biomaterial then served as a delivery vehicle, to allow surgical handling, exchange of nutrition across the transplant, and a controlled degradation. The biomaterial consisted of two outer layers of collagen and a core of a mechanically stable and slowly degradable polymer. The minced epithelium was incorporated into one of the collagen layers before transplantation. By mincing the epithelial tissue into small pieces, the pieces could be spread and thereby the propagation of cells was stimulated. After the initial take of the transplants, cell expansion and reorganization would take place and extracellular matrix mature to allow ingrowth of capillaries and nerves and further maturation of the extracellular matrix. The technique minimizes ex vivo manipulations and allow cell harvesting, preparation of autograft, and transplantation to the patient as a simple one-stage intervention. In the future, tissue expansion could be initiated around a 3D mold inside the body itself, according to the specific needs of the patient. Additionally, the technique could be performed in an ordinary surgical setting without the need for sophisticated cell culturing facilities.

Tissue engineering often includes in vitro expansion in order to create autografts for tissue regeneration. In this study a method for tissue expansion, regeneration, and reconstruction in vivo was developed in order to minimize the processing of cells and biological materials outside the body.

Conventional techniques for cell expansion and transplantation of autologous cells for tissue engineering purposes can take place in specially equipped ...

Bioengineering

Murine Full-thickness Skin Transplantation

Murine full-thickness skin transplantation is a well-established in vivo model to study alloimmune response and graft rejection. Despite its limited application to humans, skin transplantation in mice has been widely employed for transplantation research. The procedure is easy to learn and perform, and it does not require delicate microsurgical techniques nor extensive training. Moreover, graft rejection in this model occurs in a very reproducible immunological reaction and is easily monitored by direct inspection and palpation. In addition, secondary skin transplantation with donor-matched or third-party skin grafts can be performed on more complex transplant models as an alternative and uncomplicated method to assess donor-specific tolerance. The complications are low and are in general limited to anesthesia overdose or respiratory distress after the procedure. Graft failure, on the other hand, occurs commonly as a result of poor preparation of the graft, incorrect positioning in the graft bed, or inappropriate placement of the bandage. In this article, we present a protocol for full-thickness skin transplantation in mice and describe the important steps necessary for a successful procedure.

Murine full-thickness skin transplantation is a well-established model to study rejection in an alloimmune setting. Here, we provide a tutorial of each step involved in performing a BALB/c--&#62;C57BL/6 full-thickness skin transplant.

Murine full-thickness skin transplantation is a well-established in vivo model to study alloimmune response and graft rejection. Despite its limited ...

Immunology and Infection

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

Vessel-sparing Excision and Primary Anastomosis

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short bulbar or posterior urethral strictures, urethroplasty by excision and primary anastomosis (EPA) is recommended. As EPA only requires the excision of the narrowed segment and the surrounding spongiofibrosis, a full-thickness transection of the corpus spongiosum, as performed in the traditional transecting EPA (tEPA), is usually unnecessary. Jordan et al. introduced the idea of a vessel-sparing approach in 2007, aiming to reduce surgical trauma, especially to the dual arterial blood supply of the urethra, and, thus, potentially reducing the risk of postoperative erectile dysfunction or glans ischemia. This approach could also be beneficial for subsequent urethral interventions such as redo urethroplasty using a free graft, in which a well-vascularized graft bed is imperative. Nevertheless, these potential benefits are only assumptions as prospective studies comparing the functional outcome of both techniques with validated questionnaires are currently lacking. Moreover, vessel-sparing EPA (vsEPA) should at least be able to provide similar surgical outcomes as tEPA. The aim of this paper is to give an elaborate, step-by-step overview of how to manage patients with isolated short bulbar or posterior urethral strictures with vsEPA. The main objective of this manuscript is to outline the surgical technique and to report the representative surgical outcome. A total of 117 patients were managed according to the described protocol. The analysis was performed on the entire patient cohort and on the bulbar (n = 91) and posterior (n = 26) vsEPA group separately. Success rates were 93.4% and 88.5% for the bulbar and posterior vsEPA, respectively. To conclude, vsEPA, as outlined in the protocol, provides excellent success rates with low complication rates for isolated short bulbar and posterior urethral strictures.

Here, we present an elaborate and efficient protocol to treat isolated short bulbar or posterior urethral strictures with vessel-sparing excision and primary anastomosis.

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short ...

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem cells can be obtained from different tissues and several methods for isolation have been described. The presented method for the isolation of adipose-derived regenerative cells (ADRCs) can be used within many therapeutic areas because the method is a general procedure and, therefore, not limited to erectile dysfunction (ED) therapy. ED is a common and serious side effect to radical prostatectomy (RP) since ED often is not well treated with conventional therapy. Using ADRC&#8217;s as treatment for ED has attracted great interest due to the initial positive results after a single injection of cells into the corpora cavernosum. The method used for the isolation of ADRC&#8217;s is a simple, automated process, that is reproducible and ensures a uniform product. Furthermore, the sterility of the isolated product is ensured because the entire process takes place in a closed system. It is important to minimize the risk of contamination and infection since the stem cells are used for injection in humans. The whole procedure can be done within 2.5-3.5 hours and does not require a classified laboratory which eliminates the need for shipping tissue to an off-site. However, the procedure has some limitations since the minimum amount of drained lipoaspirate for the isolation device to function is 100 g.

Precise disclosure of methods and protocols is crucial for large scale uptake of stem cell therapies. Here, we present a protocol to isolate adipose-derived regenerative cells, used for a single intracavernous injection as treatment of erectile dysfunction (ED) following radical prostatectomy (RP).

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem ...

Accessing Collagen Scaffold for Single-Stage Urinary Diversion Surgery in Pigs

Surgical Model for Single-Staged Tissue-Engineered Urothelial Tubes in Minipigs

Reconstructive surgeries are often challenged by a lack of grafting tissue. In the treatment of urogenital malformations, the conventional solution has been harvesting gastrointestinal tissue for non-orthotopic reconstruction due to its abundance to reestablish normal function in the patient. The clinical outcomes after rearranging native tissues within the body are often associated with significant morbidity; thus, tissue engineering holds specific potential within this field of surgery. Despite substantial advances, tissue-engineered scaffolds have not yet been established as a valid surgical treatment alternative, mainly due to the costly and complex requirements of materials, production, and implantation. In this protocol, we present a simple and accessible collagen-based tubular scaffold embedded with autologous organ-specific tissue particles, designed as a conduit for urinary diversion. The scaffold is constructed during the primary surgical procedure, comprises commonly available surgical materials, and requires conventional surgical skills. Secondly, the protocol describes an animal model designed to evaluate the short-term in vivo outcomes post-implantation, with the possibility of additional variations to the procedure. This publication aims to demonstrate the procedure step-by-step, with special attention to the use of autologous tissue and a tubular form.

Tissue-engineered implants for reconstructive surgery rarely progress beyond preclinical trials due to laborious ex vivo culturing, which includes complex and expensive scaffold components. Here, we present a single-staged procedure designed for urinary diversion with an accessible collagen-based tubular scaffold containing autologous micrografts.

Reconstructive surgeries are often challenged by a lack of grafting tissue. In the treatment of urogenital malformations, the conventional solution has ...

Urethral Stricture Induction Followed by Buccal Mucosa Graft Urethroplasty in a Rat Model

Urethral reconstruction is an important area of expertise for urologists. The buccal mucosa is considered the best option when urethral grafting is necessary, although in some cases, it is inappropriate or needs to be optimized to repair a given stricture. Therefore, developing innovative procedures and evaluating their putative success in experimental models is crucial to fit the clinical need. With this goal, this study describes a protocol in which urethral stricture was induced by electrocautery in Wistar rats. Urethral reconstruction was performed 1 week later with a buccal mucosa graft, harvested from the lower lip, and placed in a ventral onlay fashion. A retrograde urethrogram showed a significant improvement in urethral diameter after urethroplasty compared to the respective value after stricture induction. Additionally, the graft placement was assessed by blood perfusion analysis using laser Doppler. As expected, a dark blue area corresponded to the non-vascularized buccal mucosa graft. This procedure can successfully simulate the normal pathophysiological process of urethral injury and tissue modulation, as well as urethral reconstruction using a buccal mucosa graft in a reproducible manner, and serve as the basis for future research based on tissue engineering or urethral grafts.

In the present protocol, a urethral stricture induction was developed in Wistar rats, followed by urethral reconstruction with a buccal mucosa graft. A retrograde urethrogram and laser Doppler assessment were performed, validating urethral reconstruction (after stricture formation) and graft placement.

Urethral reconstruction is an important area of expertise for urologists. The buccal mucosa is considered the best option when urethral grafting is ...