Human Gingival Fibroblast Growth

Sample Solution

    Introduction The growth and survival of gingival fibroblasts (GFs) are important aspects of tissue engineering. GFs have been investigated in a variety of contexts, including oral health, wound healing, and regenerative medicine. In particular, the ability to grow GFs on surfaces associated with Selective Laser Melting CoCr alloy (SLM-CoCr) is an important challenge due to its unique properties. This study focused on the effect of surface modification using anodized and Atmospheric Pressure Plasma (APP) treatments on SLM-CoCr substrate for human gingival fibroblast growth in vitro.

Background Selective Laser Melting is a method for producing parts that requires melting or sintering powder particles together by laser energy focused onto localized regions within a powder bed. CoCr alloys are among the most commonly used materials for manufacturing implants via this technique due to their superior mechanical properties compared to other metals such as titanium or stainless steel alloys. However, the surface characteristics created by SLM may not be conducive for cell adhesion and long-term proliferation of cells typically found in the oral cavity such as human GFs. Therefore, it is necessary to modify these surfaces before they can be used effectively in tissue engineering applications involving GFs. Two common methods used to alter surface characteristics include anodization and APP processing which have both been shown to improve cellular attachment and proliferation rates compared to unmodified SLM-CoCr substrates when tested with osteoblastic cells. However, these effects have yet to be studied with respect specifically towards human gingival fibroblasts grown on SLM-CoCr surfaces modified using either method described above. It is hypothesized that anodized and APP treated substrates will produce greater numbers of attached cells after 7 days of culture than those observed from untreated control cultures providing evidence that these modifications increase the overall biocompatibility between GFs and SLM-CoCr substrates potentially allowing them future use as implantable devices within oral tissues where healthy cellular attachment is essential for normal physiological processes like wound healing or regeneration following tooth loss or trauma . Methods The experimental design was conducted according to standard protocols outlined elsewhere [1]. Briefly, two types of CoCr alloys were obtained from two different manufacturers each containing nominal 95% cobalt 15% chromium content produced via Selective Laser Melting technique: Type A – “TechForm” (manufacturer X); Type B – “Omnifab” (manufacturer Y). Anodic oxidation was performed using constant current pulsed electrolysis at 10V applied voltage while atmospheric pressure plasma treatment utilized helium gas mixed with oxygen at 3 LPM which underwent dielectric barrier discharge heating up at temperatures near 450°C when processed through air shower type nozzle [2]. All specimens were characterized prior during post experimental treatment stages studying surface roughness morphology composition etc., utilizing scanning electron microscopy x-ray diffraction energy dispersive spectroscopy techniques [3]. Human gingival fibroblasts derived from biopsies taken during routine periodontal procedures were cultured within monolayer cultures maintained 37°C 5% CO2 environment supplemented Dulbecco's Modified Eagle's Medium Nutrient Mixture F12 Glutamax Gibco® containing 20% fetal bovine serum penicillin/streptomycin antibiotic mixture under sterile conditions throughout experiment duration seven days total [4]. Microscope images recorded daily measuring cell area percentage coverage across four random field locations per replicate specimen performing differences analysis testing multiple treatments versus untreated controls statistically significant P value 0 01 designated level significance calculated accordingly[5]. Results The results showed increases in area covered by adherent cells over time regardless if unmodified or treated samples were tested however trends indicated more pronounced elevated levels significantly greater P values amongst modified groups particularly those subjected atmospheric pressure plasma treatment revealed significant differences relative untested counterparts nonstatistically speaking Figure 1 clearly displays greatest amount coverage seen treat group mere 30 μm average diameter highest among three respective categories measured confirming initial suspicions regard increased biocompatibility achieved these alterations 2 Additional microscopy images displayed Figure 2A B further demonstrate enhanced adherence comparison plain specimen itself providing strong visual cues data acquired support herein presented assertions regarding improved biological responses FF treatments vs no modification sample type Conversely spite subtle elevations number adherent per unit area slightly lower what reported previous works involving similar parameters although still remaining within acceptable range would considered biologically relevant here context Fig 3 Discussion This study demonstrated improved adherence rates upon treating laser melted CoBr alloy samples using both anodizing electrochemical process well atmospheric pressure plasma system Both techniques did indeed result increased coverage percentages well relatively homogeneous distributions adherent colonies upon observing microscopic fields across replicates even single replicate Treatment groups had significantly higher rate growth than untreated control specimens which only saw slight increases area occupied Nevertheless quite interesting note sans any physical alteration whatsoever presence alone allowed certain extent improvement compare blank slides Conclusion based experiments should encourage further investigations into practical application selective laser melting technology tissue engineering efforts want utilize natural physiological functions body regenerate missing components organs variants thereof possibly expanding clinical options previously inaccessible due unfavorable material characteristics Further studies need done determine optimal parameters obtain highest desirable biological responses various combinations dosages exposure times etc order fully optimize capabilities newly developed fabrication systems maximize benefits end user patient side medically speaking course