Not sure if this will help, but I found this for you bro and it looks like using IGF and TGF-b can help out. Hopefully you can follow this....
Several naturally occurring polypeptide growth factors play an important role in cartilage homeostasis.
The differentiating and matrix anabolic activity of insulin-like growth factor-I (IGF-I) and transforming growth factor-b (TGF-b) are particularly
important in counteracting the degradatory and catabolic activities of cytokines, serine proteases, and neutral metalloproteases.
The manipulation of this balance in disease conditions such as
arthritis and acute cartilage injury may be possible by exogenous administration of IGF-I and TGF
The effects of these and other growth factors have been studied extensively in culture systems of many types. Most data have been generated from chondrocyte monolayer and cartilage explant cultures, where IGF-I and TGF-b generally result in elevated matrix molecule laboration, concurrent with minor to moderate mitogenic effects.
Similar results were evident in monolayer cultures of equine chondrocytes, where dose-dependent stimulation of proteoglycan production occurred in serum-free and serum-supplemented cultures.
Three-dimensional culture assessment of the effect of these same growth factors on equine chondrocyte metabolism have also been performed with fibrin gels, which provide a stable suspension culture resembling the cartilage matrix environment.
The dose-response to growth factors was largely influenced by the presence of fetal calf serum in the media. When cultured without serum supplements, these two growth factors stimulated matrix component elaboration in a dose-dependent manner; the most profound effects occurred at the highest concentrations of IGF-I and TGF-b.
Serum-free cultures are not representative of the in-vivo environment experienced by cartilage, particularly in immature horses; therefore, inclusion of serum or defined media-derived supplements are important components of culture experiments.
Enhanced proteoglycan and collagen synthesis, as well as stimulated cellular replication, were evident only at lower dose rates of IGF-I (10 ng/ml) and TGF-
b (1 ng/ml) in serum-supplemented cultures.
Increased concentrations of IGF-I (50 and 100 ng/ml) resulted in minimal further improvements. Increased levels of TGF-
b (5 and 10 ng/ml) eventually suppressed matrix synthesis and cell division.
Other studies in the horse largely focused on IGF-I because it was not detrimental to chondro-cyte metabolism when present in excess concentrations.
Several cartilage explant studies that used both normal and interleukin-1–depleted cartilage also revealed that IGF-I had a positive effect on
equine cartilage homeostasis.
Given these results, IGF-I was selected as a suitable growth factor for in-vivo studies in the horse. Other investigators evaluated articular repair following TGF-b administration.
However, synovitis and osteophyte development have been alarming features of TGF-b use in these animal studies.
Slow-release delivery of IGF-I within a cartilage remain for a minimum of 3 weeks following an initial loading dose of 20 mg. Although the dissolution rate of fibrin in the synovial environment is not known, it is not expected to vary considerably from the buffered polyionic saline used in the in-vitro elution studies. In-vivo evaluation of a self-polymerizing fibrin vehicle that is devoid of cells but is loaded with 25 mg IGF-I and injected into cartilage lesions in the femoropatellar joints showed improved cell population with more cartilage-like architecture after 6 months.
However, markers of hyaline cartilage such as type II collagen increased
to 47%, far short of the 90% minimum evident in normal articular cartilage.
Nevertheless, simple fibrin vehicle grafts used in control stifles did not significantly enhance healing. There was a mean collagen type II content of 39%, which is similar to healing in empty full-thickness defects.
Other studies using injected combinations of IGF-I and pentosan polysulfate showed attenuation of the symptoms of synovitis in OA models in sheep.
In general, IGF-I seems to have better application in combination with chondrocyte or mesenchymal stem cell grafts, where more complete cartilage repair develops.
Healing evaluation at 8 months, following implantation of a mixture of chondrocytes and 25 mg IGF-I in stifle defects of 8 horses, showed a considerably improved joint surface. There was 58% type II collagen and better neocartilage integration at the defect edges. Studies of IGF-I and
BMP-7 gene-enhanced chondrocyte function in similar transplant models suggest that both may stimulate healing beyond that seen in unstimulated
Clinical resurfacing trials in horses have used a regimen of autogenous fibrin laden with 50 mg IGF-I and 30 million chondrocytes per milliliter of fibrin.
The chondrocytes were mixed with fibrinogen and IGF-I with activated thrombin to provide a 2-component system for immediate injection.The polymerization process developed immediately upon injection into the articular defect. Currently, the predominant application has been OCD and subchondral cystic defects of the fetlock (8 horses) and stifle (43 horses), although several shoulder and carpal articular lesions have been treated. The fibrin polymer and growth factor have been well tolerated
and have effected resolution of effusion and lameness.
Assessment of stifle OC cyst cases 1–5 years after implant indicates that the horses respond by increased bone deposition in the subchondral plate
and then generally throughout the cyst over the ensuing first year.
Soundness has taken as long as 1 year to develop, but 22 of 30 horses beyond the first post-operative year have remained in active work without lameness. Six of 8 horses with grafted fetlock subchondral cysts have been evaluated beyond 1 year and all returned to racing or nonracing athletic work.
Several members of the bone-morphogenetic protein (BMP) family also have considerable benefits in cartilage healing. Of these, BMP2 and BMP7 show the most promise. In-vitro studies show that BMP2 has matrix stimulatory effects similar to IGF-I. Long-term in-vivo studies show enhanced cartilage repair in rabbits.