Adhesion of MC3T3‐E1 cells bound to dentin phosphoprotein specifically bound to collagen type I

Abstract

Dentin sialophosphoprotein (DSPP) is a member of the SIBLING (small integrin binding N-linked glycoprotein) family of proteins commonly found in mineralized tissues. Dentin phosphoprotein (DPP) is a naturally occurring subdomain of DSPP that contains the cell binding RGD sequence. Previously, the orientation and conformation of other SIBLING family members specifically bound to collagen I have been investigated with respect to their cell adhesion properties. In this study, the orientation of DPP under similar circumstances is examined, and the results are discussed relative to the previous investigations. Radiolabeled adsorption isotherms were developed for DPP adsorbing to both tissue culture polystyrene (TCPS) and collagen coated TCPS. Then, a MC3T3-E1 cell adhesion assay was performed on TCPS and collagen coated TCPS in the presence of identical amounts of adsorbed DPP. It was discovered that there was a significant difference in the number of bound cells on the TCPS and collagen coated TCPS, with a preference for TCPS. Furthermore, a cell inhibition assay was conducted to confirm that the cell binding that occurred was due to specific integrin interactions with the RGD sequence of DPP. These results suggest that the orientation of DPP, rather than its conformation, dictates the accessibility of the cell binding RGD domains of DPP and that the RGD sequence in DPP is less accessible when DPP is specifically bound to collagen. The results obtained in this study are in stark contrast to previous studies with related SIBLING proteins, and suggest that DPP does not play a key role in cell binding to the collagen matrix of developing bone.

Enhancement and mechanisms of MC3T3‐E1 osteoblast‐like cell adhesion to albumin through calcium exposure

“…In order to screen the bioactivity of Ca‐modified BSA, a high cell seeding density (1 × 10 5 cells/mL) adhesion and viability study was conducted because the high seeding density facilitates greater adhesion levels that allows for trends in the cell viability to be clearly identified. Adhesion was evaluated after 2 H of cell exposure, while viability was assessed following 2 H of adhesion, as well as after removal of the cell solution and replacement with low supplemented media (1% FBS) and an additional 22 H of culture, based on similar studies in the literature investigating protein‐cell interactions at biomaterial interfaces 5,7,8,24,25 . Additionally, to maximize the impact of calcium on the conformation of albumin, cell adhesion was first evaluated immediately following the adsorption process without any surface rinses.…”

“…Two different inhibitors were used in an effort to identify integrin‐mediated binding interactions. A GRGDSP peptide was used to inhibit RGD‐binding integrins using a peptide concentration taken from prior studies 24,25 . BTT 3033 was used to inhibit integrin–collagen‐binding interactions that typically involve the GFOGER sequence in collagen.…”

Bone Regeneration Induced by Strontium Folate Loaded Biohybrid Scaffolds

“…Bone tissue consists mainly of the protein collagen (~90%), and of cells with a role in bone formation and growth, repair and synthesis of the extra cellular matrix [23,24]. The matrix is formed by a small binding integrin-linked to glycoprotein N. The sibiling protein family is formed by five members: (a) osteopontin (OPN), (b) fosfoglycoprotein of the extracellular matrix (MEPE), (c) bone sialoprotein (BSP), (d) dentin matrix protein 1 (DMP1), and (e) dentin sialophosphoroprotein (DSPP).…”

“…Undoubtedly, these proteins have an important role in bone development, i.e., facilitating cell adhesion, nucleation, and mineral maturation. Nonetheless, researchers are still trying to elucidate the exact role of each protein in natural bone development [24,25].…”

Anti-Microbial and Anti-Wrinkle Effect of Kaempferol and Kaempferol Rhamnosides isolated from Hibiscus cannabinus L.

“…피부의 기질 단백질은 피부 에 강도와 장력을 주는 콜라겐이 90% 이상 함유되어 있는데 type Ⅰ collagen이 대부분이고 type Ⅲ collagen이 소량 포함 되어 있으며 피부에 탄력을 주는 탄력섬유가 3~4% 정도 함 유되어 있다. 콜라겐은 섬유아세포와 작용에 의해 합성되며 콜 라겐 분해효소의 작용에 의해 분해되므로 세포외 기질 단백질 의 결핍을 막아서 피부 주름을 예방하기 위해서는 교원질 합 성을 증가시키고 교원질 분해효소의 작용을 억제시키는 것이 필요하다 (Zurick et al, 2012). 이처럼 피부노화에 대한 연 구는 콜라겐의 양과 콜라젠을 분해하는 효소 (Matrix metalloproteinases; MMPS)의 생합성을 억제하거나 활성을 저해하는 방식 혹은 항산화, 항노화 물질을 이용하여 피부 세포 를 보호하려는 노력이 이루어지고 있다 (Neena et al, 2011).…”