The Porosity and Human Gingival Cells Attachment of Synthetic Coral Scaffold for Bone Regeneration

Porosity and interconnectivity play an important role in the success of tissue engineering because it affects cells to live and grow. Coral has been used as a bone replacement material because the structures resemble bone and have mechanical bone properties. In this study, the synthetic coral scaffold was developed to mimic the natural coral. This study aims to investigate the porosity of the scaffold and its biocompatibility while it is attached to human gingival cells. Synthetic coral scaffold in various compositions were prepared, porosity percentage measurement and human gingival cell attachment test were done. An optimum ratio of the scaffold with gelatin: CaCO3, having the highest porosity and cell attachment is obtained in 5:5. The result of this study presented that synthetic coral scaffold could provide the microenvironment to cells for life because it is supported by the highest percentage of porosity.

Characteristic of Synthetic Coral Scaffold for Cell Environment

Introduction. Synthetic coral scaffold is fabricated the mimicking of natural sea coral as a scaffold for bone regeneration [1]. Scaffold is performing functions as a micro environment for cells attachment, growth, proliferates, differentiates until it can form new bone tissue. The proper design is needed to produce the scaffold [2]. The purpose of this study was to investigate the characteristics of synthetic coral scaffold for micro environment of cells by observing cell attachment, hydrophobicity, and scaffold porosity. Experimental. Synthetic coral scaffold consists of bovine gelatin dan CaCO3 by weight, the concentration that be used are 4:6. Sodium citrate is used as dispersant. Thick film like scaffold was prepared for this study [1,3]. Vero cell line was used for observing cell attachment to investigate the biocompatibility the scaffold. The hydrophobicity was observed with distilled water droplets dripped on the scaffold surface, be analyzed in a photograph taken by the camera and then measured the angle. Percentage of porosity was measured using Archimedes law in absolute ethanol. Results and Discussion. Vero cells attached successfully into scaffold. Cell viability percentage is 91,77 % from the absorbance value of the MTT assay. It presented that the scaffold has biocompatibility character. However, the percentage of porosity is 55,85%, so the scaffold has enough porosity for cell attachment. Porosity serves for the diffusion of nutrients, gases and removes the residual metabolism resulting from cell activity that has grown on scaffold. The good porosity value of the scaffold is 50-90%. The higher the porosity value the better the scaffold. Hydrophobicity scaffold appears from the contact angle of 81.4°, the cohesion is greater than the adhesion. This shows the greater synthetic coral scaffold hydrophobicity, which is affected by surface roughness from scaffold porosity. The greater hydrophobicity will also prolong the degradation of the scaffold, thereby enabling cells to proliferate, differentiate and produce bone matrix. Conclusions. Synthetic coral scaffold provides the micro environment for cell, high hydrophobicity allows longer degradation for proliferation and differentiation of bone cells, and porosity that allows cells to be inserted within the scaffold.

Degradation profile of synthetic coral scaffold in cell culture media

<p>The scaffold is one of the factors in tissue engineering that determine the success of bone regeneration. The important characteristic of the scaffold is able to degrade gradually. In vitro study using cells, the scaffold will be exposed to culture media. Therefore, degradation profile for scaffold needs to be examined. This study aims to investigate the degradation profile of synthetic coral scaffold in cell culture media using pH measurement. The method used the synthetic coral scaffolds were prepared from denaturalized collagen (gelatin) and calcium carbonate (calcite) with a concentration of 5:5 and 4:6 weight % in aqua dest. The scaffold<span lang="IN" style="">s</span> were fabricated in membrane thick film which was then physically crosslinked.&nbsp; <span lang="IN" style="">The </span>10 % <span lang="IN" style="">of gelatin scaffold was used as a control.</span>&nbsp; The scaffolds were incubated in cell culture media (non-phenol red Dulbecco’s Modified Eagle Medium) for 1 until 8 days, and pH changes of the medium were measured. As the result, Profile of degradation on day 1 to day 4 showed the 5:5 scaffold had the smallest degradation. The results indicated the significant different between scaffold concentration in day 1 (p=0.005), 5th(p=0.03), and 6th day (p=0.011). At the end of incubated days, the pH changed but not significantly different. LSD showed the significant differences between scaffold (5:5 and 4:6) with control and no significant difference between 2 concentrations of the scaffold. The conclusion of this study is the synthetic coral scaffold degraded gradually until the end incubation time and between concentration had different degradation profile in the early incubation time using pH measurement.<br></p>

Human Mesenchymal Stem Cells Behavior on Synthetic Coral Scaffold

For bone tissue engineering, corals have long history to be used as scaffold to promote bone regeneration. However the use of a lot of corals may damage their habitates. For this reason, a strategy to mimic coral in a synthetic form is needed. As an ideal scaffold, synthetic coral must provide structure and initial support for cell attachment and proliferation. The aim of this study was to investigate the attachment and proliferation of human Mesenchymal Stem Cells (h-MSC) on synthetic coral scaffold, to provide information on the behavior of h-MSC on the designated scaffold. Synthetic coral scaffolds were prepared from bovine gelatine and CaCO3 with 5:5 in 10% w/v concentration in aquadest. Sodium citrate was used as dispersant in the suspension. Gelatin-CaCO3 suspension was moulded in a plastic cover of 24 well plate, then freezed at -20°C for 24 hours, freeze dried for 24 hours and continued by dehydrothermal crosslinking for 72 hours. After the fabrication, synthetic coral scaffolds were subjects to cover the bottom of the well for cell culture. Human Mesenchymal Stem Cells (h-MSC) were seeded and divided into 2 groups, control group without scaffold and the one with scaffold. All groups were incubated for 3, 6, and 24 hours. Cells attatchment were determined by deduction of the cells unattached from total cells seeding. Proliferation of h-MSC were done in 3 groups ie., control group without scaffold, scaffold only and scaffold incorporated Platelet Rich Plasma (PRP) in the bottom of well. All groups were incubated for 24, 48 and 72 hours. Human Mesenchymal Stem Cells attached faster to synthetic coral scaffold than the control. Its proliferation behavior was faster in the scaffold incorporated PRP, showing better interaction of scaffold and cells with the incorporation of morphogenetic factor.