Video abstract of an original research “Mucoadhesive microspheres of Maraviroc and Tenofovir designed for pre -exposure prophylaxis of HIV-1: an in-vitro assessment of the effect on the vaginal lactic acid bacteria microflora” published in the open access journal HIV/AIDS – Research and Palliative Care by Sabdat Ekama, Margaret Ilomuanya, Chukwuemeka Azubuike et al.
My name is sabda akama and i’ll be presenting this video abstract on behalf of the co-authors on the article titled mocha adhesive microspheres of maravirak and tenofova designed for pre-exposure prophylactics of hiv-1 and in vitro assessment of the effect on vaginal lactic acid bacteria microflora heterosexual intercourse is a major mode of male to female
Transmission of hiv-1 infection and women account for a greater percentage of hiv prevalence globally this has necessitated the development of pre-exposure prophylaxis measures such as microbicides as prevention strategies previous microbicide formulations had challenges to his use such as short resident time the vagina leakage and coital dependent use this has led
Researchers to the development of targeted drug delivery systems such as microspheres that can target the drug to the site of action and improve vaginal resident time this study aims to formulate and evaluate microspheres of the antiretroviral drugs maraviroc and tenofova intended for a candidate vagina microbicide and assess its effect on the vagina lactic acid
Bacteria microflora a full factorial experimental design at two levels of concentration requiring tool to the power k runs was used to determine the varying effect of independent factors such as concentration of the polymer the cross-linking agent and curing time on dependent variables such as particle size much adhesion and encapsulation efficiency maravi rock
And tenofova microspace were prepared using ionic gelation technique in which the polymers containing the anti-java drugs were cross-linked with sodium tripolyphosphate to form micro particle carriers for the drugs varying weights of kyto sun were dissolved in 10 mil of 4 percent acetic acid instead using a magnetic stereo for 30 minutes with subsequent addition
Of 10 milligram of the individual antiviral drugs the resultant dispersion was collected in a syringe and added drop wise into a solution of sodium tripoli tri-polyphosphate to achieve cross-linking the jelly beads formed we are allowed to stand for a curing time of 15 to 30 minutes which is the wet microscope that we can see and subsequently it was decanted
Washed with milky water and dried at 45 degree for 12 hours which is the dried microspace that we can see and the microspheres were viewed from the microscope as seen in plate 1b mocha adhesion test using sheep vagina tissue was used to evaluate the muco-adhesive ability of the formulated microspace sheet vagina tissue was obtained from a freshly slaughtered
Sheep in an aperture the vagina tissue was cut into the size of a glass slide and raised with prepared simulated vagina fluid and attached to a glass slide the microspheres were counted and 30 pieces were attached to the vagina tissue which was subsequently immersed into a beaker containing 900 ml of the stimulated vagina fluid the slide was attached to the
Arm of a disintegration apparatus and given a vertical reciprocating movement for approximately 30 cycles per minute at the end of one hour and at the end of 12 hours the number of microspheres that were still attached to the tissue were counted and the percentage much adhesion was calculated by expressing the value as a percentage of the originally attached
Number of microspheres drug release studies was carried out according to the method of agro holly at all in which the formulated microspheres where each was suspended in three mil of a mixture of seminar flue simulant and vagina flu similar and the dispersion was transferred into a 10 mil capacity dialysis membrane the membrane was maintained in a 20 main release
Medium contained in a dialysis tube and the whole system was placed in the conical flask then placed in the shaking water but and the temperature was maintained at 37 degrees at periodic time intervals of zero hour one hour three hours all through to 72 hours one minute sample was withdrawn from the release medium in the dialysis tube for hplc analysis and
At each point of withdrawal an equivalent volume of the release medium was used to replace the one that was redrawn to maintain sync condition the amount of maravirock and tenofova released was determined using hplc analysis an ideal microbicide should not disrupt the activities of vaginal lactic acid bacteria which helps to maintain the protective acidic ph
Of the vaginal environment lactobacillus acid was therefore carried out to evaluate the effect of the microspace on isolated vaginal lactic acid bacteria atypical approval was obtained to conduct this study and it was carried out according to the declaration of helsinki the women gave consent to participate in the study and vagina samples were collected using
Stereo vaginal swab sticks and the lab organisms were isolated on mrs aga dna extraction was carried out using the gina bioscience dna extraction kit and polymerase chain reaction was carried out using the lab-specific primers these primers produced an application size of 526 base pair by all lactic acid bacteria the amplified pcr products were purified using
The general bioscience purification kit and it was assessed for the it was sequenced for the 16s rna gene and the corresponding sequences were identified using the online blast search and submitted to the gene bank for gene essential number assignment the identified organisms were lactobacillus fermentum and enterococcus fecalis an nts acid was carried out in
Which the organisms were exposed to the microsphere suspension for 48 hours after which a microplate reader was used to determine the absorbance and the percentage viability which is a measure of the number of life cells was determined hiv efficacy testing was evaluated against the hiv-1 bal viral stream these are the mbl cells were plated into 96 well plates
And 100 microliter of serial dilutions of maravirak and tenofova microspheres were added to the wells non-ordinal nine gel was used as the negative control and the plates were incubated for 48 hours after with the medium was removed and replaced with hundred microliter of bright glow and the luminescence was measured after which a dose response curve was used
To describe hiv infectivity rate the results as follows table 1 shows the predicted and experimental values of the optimal conditions of microspace obtained by numerical optimization numerical optimization was used to identify the conditions for the optimized formulation and a prediction of the outcome variables the two sets of values shows adequate proximity
Between the experimental and the predicted values the optimal formulation had moderate particle sizes high percentage nuclear adhesion and good encapsulation efficiency an analysis of the in vitro drug release for marijuana and tenofovir from the microspheres was carried out by fitting the release data into the various kinetic models to determine the coefficient of
Correlation the model with the highest coefficient of correlation best explains a drug kinetic release pattern maravirock release followed a zero order kinetic model and a super case through drug transport mechanism while tenofova release followed a hikuchi model release kinetics and a non-fiction drug transport mechanism results from the lactobacillus viability
Assay shows that lactobacillus fermentum after a 48 hour exposure to the microsphere suspension had a percentage viability of 93.9 percent while entire cocoa’s facalis had a percentage viability of 89.7 percent there was no statistically significant difference between the absorbers of l fermentum and the negative control and the absorbance of ethicalis and negative
Control but there was a statistically significant difference between the absorbance of the organisms and the positive control and this shows that the organisms were able to thrive in the presence of the microspheres the pre-exposure prophylactic ability of the combined optima maraviroc and tenofova microspheres were evaluated via an in vitro concentration response
Procedure using anodizing online as control the hiv infectivity of the combined microspheres compared with neglect online showed a marked difference in hiv infectivity rate against the hiv-1bl viral strain the observed continuous decline in hiv infectivity of the microspheres indicates adequate release of the drugs from the polymer and anti-hiv activity in
Conclusion the anti-java drugs loaded in the microscopes had good mocha adhesion which is a potential for prolonged residence time in the vagina the anti-chiva drugs were adequately released from the microspace and showed efficacy against the hiv-1ba of viral strain there was no significant disruption in the growth of lactic acid bacteria which constitute the
Valid bacteria microflora of the vagina thank you all for your attention this project is supported by the fogarty international center of the national institute of thought under award number d43tw010934 the content is solely the responsibility of the authors and does not necessarily represent the official views of the national institute of health thank you
Transcribed from video
Maraviroc and Tenofovir for pre-exposure prophylaxis of HIV-1 – Video abstract [ID 291065] By Dove Medical Press