This method has a potential to be utilized as a new clinical strategy for knee osteoarthritis. This technique is safe, simple for local injections, and the treatment is effective in animal experimentation. Silver nanoparticles might be applicable in other inflammation disease and it needs further research.
It is important then to be careful when injecting the knee size of the mice, ensuring the correct procedure. Assessing the hand fill of the knee joint at the knees and avoid any drug leakage. Begin by placing an anesthetized mouse on a surgical table.
Tape the limbs of the mouse, then shave the fur with a razor. Next, disinfect the shaved region thrice with alternating scrubs of 0.1%iodoform and alcohol. Using scissors, make an incision on the skin, sequentially exposing the subcutaneous tissue and the infra patella ligament.
With a one milliliter insulin syringe, inject 10 units of type two collagenase into the joint cavity. Suture the subcutaneous tissue, then suture the skin. Now sterilize the sutured area with 0.1%iodoform.
Place the mouse into individually ventilated cages once they wake up from the anesthesia. Start by adding 400 microliters of type one collagen to a 1.5 milliliter micro centrifuge tube placed on ice. Now, add 200 microliters of PBX, mix well and place the tubes on ice.
Add 400 microliters of silver nanoparticles to this suspension, making the final nanoparticle concentration to one milli mole. Mix the suspension well. One week after the collagenase injection, remove the KOA mice from their cages.
Before injecting silver nanoparticles, anesthetize and surgically prepare the mouse as previously demonstrated. Use scissors to sequentially expose the skin, subcutaneous tissue and knee ligaments. Then, using the needle of a one milliliter insulin syringe, enter the joint cavity at a 15 degree angle and slowly inject 20 microliters of the silver nanoparticle collagen mixture.
During the injection, the knee joint will swell, ensuring a successful drug injection. After suturing and sterilizing the surgical site as earlier, place the mice into individually ventilated cages once the anesthesia wears off. Surgically sterilize and dissect the limbs of the euthanized mouse.
Harvest the knee joints including the femur and tibia, and remove the muscle tissue. Collect the knee joint tissues, including the femur and tibia and the surrounding soft ligament. Encapsule in 10%formalin for preservation and fixation.
After overnight fixation of the tissues, embed the sections in paraffin. Using a microtome, cut the embedded tissues into 0.4 micro thick sections. Subject the tissues to different staining techniques.
The average body weight of the KOA mice was significantly lower than that of the mice in the control group. The average body weight of the mice in the type two collagenase and silver nanoparticle group was higher compared to the KOA mice. HE staining showed that the synovial thickness of the mice in the KOA group was significantly higher compared with the control group.
The synovial thickness of the group treated with nanoparticles was reduced relative to the KOA group. Vascular hyperplasia of the synovium was observed in the KOA group and was found to be significantly reduced in the nanoparticle treated group. Safranin-O staining revealed a destroyed cartilage matrix in KOA mice, while the mice treated with the nanoparticle collagen mixture showed a significantly better cartilage matrix.
Morphological feature scores were higher in the collagen II treated group relative to the control group. Immunohistochemical staining demonstrated reduced neutrophil infiltration in the synovial regions of the nanoparticle treated mice as compared to the KOA group. It is important to avoid drug blockage during the injection of type II collagenase and several nanoparticles.
The therapeutic effect of several nanoparticles could be attempted on animal models of anterior cruciate ligament injury.
