Bovine Collagen Peptides Compounds Promote the Proliferation and Differentiation of MC3T3-E1 Pre-Osteoblasts

Abstract

ObjectiveCollagen peptides (CP) compounds, as bone health supplements, are known to play a role in the treatment of osteoporosis. However, the molecular mechanisms of this process remain unclear. This study aimed to investigate the effects of bovine CP compounds on the proliferation and differentiation of MC3T3-E1 cells.MethodsMouse pre-osteoblast cell line MC3T3-E1 subclone 4 cells were treated with bovine CP compounds. Cell proliferation was analyzed by MTT assays and the cell cycle was evaluated by flow cytometry scanning. Furthermore, MC3T3-E1 cell differentiation was analyzed at the RNA level by real-time PCR and at the protein level by western blot analysis for runt-related transcription factor 2 (Runx2), a colorimetric p-nitrophenyl phosphate assay for alkaline phosphatase (ALP), and ELISA for osteocalcin (OC). Finally, alizarin red staining for mineralization was measured using Image Software Pro Plus 6.0.ResultsCell proliferation was very efficient after treatment with different concentrations of bovine CP compounds, and the best concentration was 3 mg/mL. Bovine CP compounds significantly increased the percentage of MC3T3-E1 cells in G2/S phase. Runx2 expression, ALP activity, and OC production were significantly increased after treatment with bovine CP compounds for 7 or 14 days. Quantitative analyses with alizarin red staining showed significantly increased mineralization of MC3T3-E1 cells after treatment with bovine CP compounds for 14 or 21 days.ConclusionsBovine CP compounds increased osteoblast proliferation, and played positive roles in osteoblast differentiation and mineralized bone matrix formation. Taking all the experiments together, our study indicates a molecular mechanism for the potential treatment of osteoarthritis and osteoporosis.

The effect of different molecular weight collagen peptides on MC3T3-E1 cells differentiation

“…The formation of osteoblast mineralization nodules reflects the ability of bone cell differentiation. Results here showed that different molecular weight collagen peptides can increase the OC level in MC3T3-E1 cells and mineralization of osteoblasts, similar as our previous results [21].…”

Collagen-derived dipeptide prolyl-hydroxyproline promotes differentiation of MC3T3-E1 osteoblastic cells

“…These results indicate that Pro-Hyp can promote the differentiation of osteoblasts by upregulating gene expression of Runx2 and Osterix. Previous reports have shown that treatment with crude CP increases the expression of the Runx2 and the Col1a1 genes in osteoblastic cells [13,14]. The regulation by Pro-Hyp in MC3T3-E1 cells would explain a mechanism of the protective effect of CP in bone metabolism.…”

“…Previous reports have shown that crude CP stimulates the proliferation of osteoblast cells. In one previous report, crude CP was used that may include various collagen-derived di-, tri-, and oligo-peptides such as Pro-Hyp, Pro-Hyp-Gly, Hyp-Gly, or Ala-Hyp [12][13][14]. It is speculated that these collagen-derived peptides, except Pro-Hyp, have the ability to enhance osteoblast proliferation.…”

“…Furthermore, some studies have indicated that crude CP has a beneficial effect on the proliferation or differentiation of osteoblastic cells [12][13][14]. Therefore, CP may also play an important role in bone remodeling.…”

Hydrolysed collagen from Lates calcarifer skin: its acute toxicity and impact on cell proliferation and collagen production of fibroblasts

“…Plinia peruviana extract having antioxidative activities was reported to demonstrate the potential for stimulation of L929 cell proliferation and wound healing properties (Pitz et al, 2016). Liu et al (2014) found that collagen peptides could promote osteoblast proliferation by inducing cell cycle arrest in G2/S phase (DNA synthesis) performed by flow cytometry scanning. In addition, VitC has been known for promoting skin health by inducing the synthesis of a normal mature collagen network by maintaining optimal lysyl and prolyl hydroxylase activity, two essential enzymes in the collagen biosynthesis pathway (Boyera et al, 1998).…”