In-Vitro study: Effects of Tendoactive® on collagen fibrillogenesis in a 3-dimensional culture of human tenocytes

Objective

To evaluate the potential effectiveness of Tendoactive® on collagen fibrillogenesis (formation of collagen fibrils) and tenomodulin* (TeM) levels in the presence/absence of IL-1ß in a 3-dimensional culture of human tenocytes.

*glycoprotein predominantly expressed in tendons and ligaments.

Methods

Primary human tenocytes in 3-dimensional high- density cultures were incubated for 0-14 days under different treatments. Tenocytes were either treated with: 1. Control / 2. Tendoactive® / 3. Negative control: IL-1ß. Pre-treatment with Tendoactive® (24 hours) + co-treatment with Tendoactive® and IL-1ß (14 days).

Transmission Electron Microscopy (TEM) was used to investigate the potential efficacy of Tendoactive® in the extracellular matrix (ECM) and the orientation, organization, and morphology of the collagen fibrils. Western Blot was used to determine if tenocytes incubated in the formulation containing Tendoactive® prevented the upregulation of catabolism induced by IL-1ß, which has been demonstrated in past research, leads to the downregulation of matrix-specific production of proteins, such as tenomodulin.

Results

The treatment with Tendoactive® counteracted the catabolic effects of IL-1ß. Cells displayed characteristics consistent with healthy tenocytes, including abundant and well-organized extracellular matrix consisting of thick fibrils of collagen. Tenocytes displayed a high amount of euchromatin indicating a high level of cellular activity and with a high rate of protein biosynthesis (Figure 1).

Western Blot demonstrated a considerable prophylactic effect of Tendoactive® on human tenocytes co-treated with IL-1ß on TeM synthesis. Additionally, an anabolic effect of the formulation was observed: cells showed higher tenomodulin levels compared to the untreated control cells (Figure 2).

Conclusion

Data suggests that Tendoactive® may enhance tendon healing by:

-Increasing the amount of collagen in the healing tendon

-Promoting better collagen reorganization

-Increasing synthesis on tenomodulin, which plays a role in tendon development

-Improving the physical and mechanical properties of the tendon.