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Novel osteogenic, antimicrobial GBR solution for peri implants tissue engineering- An in vitro study.
EAO Online Library. Y Nelogi S. Oct 9, 2018; 232458
Santosh Y Nelogi
Santosh Y Nelogi
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Guided bone regeneration(GBR)is a therapeutic procedure for bone and peri-implant defects. The conventional membranes possess many bio-functional, mechanical, structural limitations, they lack osteogenic and bactericidal potential.In this research Cissus quadrangularis(CQ),hyloronic acid(HA)and chitosan(CH)composite polyelectrolyte solution was prepared. CH-has osteoconductive properties.CQ has high contents of Ca++, b-sitosterol d-amyron makes it osteogenic.HA with CH forms polyelectrolyte compwe hypothesized that the next-generation (GTR GBR) membranes will be a biologically active, antimicrobial and functionally in liquid form closely mimic the native ECM. Aim- To develop bioresorbable, bioactive and antimicrobial GBR solution which is thermo reversible in nature nature for the expression of enhanced bone tissue engineering.Preparation of CH-HA- CH (3gm in 100 ml) was dissolved in diluted acetic acid,HA was added with constant stirring. CQ Extract was prepared (100gmin 500ml).(CH- HA- CQ)solution with cross linking agent was prepared by mixing in 50-50 by weight. Micro particles of (CH- HA- CQ)were prepared by dry spray technique,prepared micro particles were added to ploximor 407 at concentration of 18% . IN VITRO ANALYSIS.The graft was distributed to cylindrical molds which was kept at a temperature of 37°C then transferred for analysis.The ECM properties were assessed by proteome analysis by mass spectrometry, and SEM.The anti microbial property was evaluated by disc diffusion, spread plate.The proliferation assay was assessed byMTTassay by at 570 nm. ALP ASSAY-was determined at 405 nm ..Matrix mineralization was assessed by von Kossa histochemical staining. All experiments were conducted in triplicate. Data are expressed as the mean standard deviation.ANOVA was conducted. (P < 0.5)The obtained hydro gel membrane was characterized by SEM and the degradation of membrane was evaluated by adding phosphate buffered solution and lysozyme over the period of 90 days. The permeability of CH CQ HA was evaluated at room temperature by coefficient of p-nitrophenol using a dialysis apparatus. The GBRsolution was stable at room temperature, but displayed sigmoidal increases at 37°C, degradation test result showed that the graft degraded by about 30–40% of initial weight after 90 days. SEM revealed good structural porosity in the gel. Proteome analysis of the ECM-solution and gel mass spectroscopy identified the most abundant proteins.The antibacterial activity of graft showed effective results against bacteria. MTT assay revealed proliferation of proliferation of osteoblast**cells (0.24 0.007) compared with control (0.22 0.007) (P < 0.34).while ALP activity of MG 63 cells had the highest value of the ALP activity (0.23 0.005) . The present work demonstrates an osteogenic injectable GBR solution which is biomimetic,angiogenic and antimicrobial in nature .This hydrogel holds the distinctive composition specific for bone ECM, After injection, the thermoresponsive behavior of the **solution allows it to polymerize and form a porous gel at body temperature.The results presented indicate that injectable ECM materials may offer a promising alternative in the treatment of bone defects .
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