Proteolytic digest derived from bovine Ligamentum Nuchae stimulates deposition of new elastin-enriched matrix in cultures and transplants of human dermal fibroblasts

Abstract

Background: Diverse topical products and injectable fillers used for correcting facial wrinkles induce rather short-lived effects because they target replacement of dermal collagen and hyaluronan, matrix components of limited biologic durability. Objective: Present studies were aimed at stimulation of fully differentiated human dermal fibroblasts to resume deposition of new extracellular matrix rich of elastin, the most durable and metabolically inert component of dermal ECM. Methods: We have created a novel proteolytic digest of bovine ligamentum nuchae (ProK-60), and tested its potential biological effect on dermal fibroblasts derived from females of different ages. Northern blots, quantitative immunohistochemistry and metabolic assays were used to assess effects of ProK-60 on proliferation and matrix production in primary cultures of dermal fibroblasts, in cultures of skin explants and after implantation of stimulated fibroblasts into the skin of athymic nude mice. Results: ProK-60 increased proliferation (25–30%) of cultured dermal fibroblasts and significantly enhanced their production of new elastic fibers (>250%) and collagen fibers (100%). These effects were mostly mediated by stimulation of cellular elastin receptor. In contrast, ProK-60 inhibited production of fibronectin (−30%) and chondroitin sulfate proteoglycans (−50%). ProK-60 also activated proliferation of dermal fibroblasts, mostly derived from the stratum basale and induced deposition of elastic fibers in cultures of skin explants. Moreover, human fibroblasts pre-treated with ProK-60 produced abundant elastic fibers after their injection into the skin of athymic nude mice. Conclusion: The described biological effects of ProK-60, including its unique elastogenic property, encourage use of this compound in cosmetic formulations stimulating rejuvenation of aged skin.