Loading

Journal of Pharmacology and Clinical Toxicology

Thermo Reversible Buffered Tenofovir Nano Gel for Vaginal Delivery

Research Article | Open Access

  • 1. Department of Pharmacy, Howard University, USA
+ Show More - Show Less
Corresponding Authors
Pradeep Karla, Department of Pharmacy, Howard University, Georgia Avenue, NW Washington, DC 20059, USA, Tel: 847-450-1842
INTRODUCTION

Tenofovir is an antiretroviral drug known as nucleotide analogue reverse transcriptase inhibitors, which block reverse transcriptase, an enzyme crucial to viral production in HIVinfected people [1]. It is currently considered the preferred first line treatment for HIV because of its potency, overall low toxicity, and convenience of dosing [2]. This drug is administered orally in the form of disoproxil ester, which is deesterified to achieve a bioavailability of more than 20% [3] This prodrug has a plasma stability issue and non-specific distribution. It is also reported that parenteral administration showed nephrotoxicity [4,5].

Literature revealed that PLGA, is a FDA-approved and widely accepted biodegradable copolymer used in nanoparticle formulation, which can also provide the sustained release of an encapsulated drug. Application of FDA approved excipients may feasible to clinical testing of final formulation. Designs of nanoparticles encapsulated drugs are still challenging area for the formulation scientists. Nanoparticles could be site specific delivery by size dependent uptake by affected cell and it may also improve drug efficacy by release the drug from the nanoparticles over a prolonged time [6]. Zhang et al. [7] studied the pH responsive PLGA/ methacrylic acid copolymer (Eudragit® S-100, or S-100) nanoparticles having TNF and results suggested the alternative controlled drug delivery system for intravaginal delivery TNF. Another formulation based on lipid were studied by Xu et al. [8] and reported that Liposomes have low encapsulation efficiency problem.

Nano size based formulation in suspension form may face the problem of low retention time at desired site that ultimately will lead to poor bioavailability. Therefore we hypothesized that if we can increase the retention of nanoparticles by gel matrix at vaginal site that may improve efficacy of TNF. This study was designed to develop thermoreversible gel containing TNF loaded PLGA nanoparticles. Final formulation will be in solution form at 2-8o C and covert into gel matrix dispersed TNF loaded PLGA nanoparticles.

MATERIALS AND METHODS

Tenofovir was purchased from Waterstonetech, LLC, USA. 75:25 poly(DL-lactide-co-glycolide) (PLGA) was purchased from DURECT, USA. Ethyl acetate, Pluronic®F-68(PEG-PPG-PEG), Poloxamer 407, Pluronic® F-108 and poly (vinyl alcohol) 98-99% hydrolyzed were purchased from Sigma-Aldrich, USA. Sodium phosphate monobasic anhydrous USP, HPLC water, HPLC grade acetonitrile were purchased from Fisher scientific, USA. Sodium phosphate dibasic heptahydrate was purchased from Affymetrix, Inc, USA. Millex® -GV, syringe driven filter unit Durapore® (PVDF) 0.22 µm membrane was purchased from Millipore, USA

Preparation of TN F loaded PLGA nanoparticles by solvent diffusion

Briefly, TNF (10-100 mg) and the polymer (100–400 mg), PLGA were co-dissolved in 5 ml DMSO. The organic phase was added drop wise (2ml/min.) to aqueous phase, containing Triton X 100, under homogenization at 4000-5000 rpm for 30 min. (Power Gen 1000, fisher scientific, USA). Suspension was ultrasonicated for 5-10 minutes and ultra-centrifuged at 15000 rpm for 90 min (Beckman L8-70 M Ultra-centrifuge, Brea, CA, USA) to collect NPs and then washed with distilled water to remove the surfactant. Settled pallet was re-suspended in distilled water and tested for particles size by DLS and SEM.

Preparation of TN F loaded PLGA nanoparticles by emulsification-solvent evaporation

TNF-loaded PLGA nanoparticles were prepared using the modified emulsion/solvent evaporation technique. PLGA (75:25) was dissolved in ethyl acetate and added to aqueous phase containing pluronic®F68 and TNF to prepare a primary emulsion and then it was ultrasonicated (Pulse: 0.9/0.1;amplitude 30- 35%) for 10 minutes on ice batch. Primary emulsion added to secondary aqueous phase (2% w/v PVA solution) and ultrasonicated for 5 minutes than ethyl acetate evaporated at room temperature by constant magnetic stirring for 12-18 h. The prepared nanoparticles were separated by ultracentrifuge and supernatant discarded and settled pellets were collected in lyophilization vial and lyophilized at -50o C for 24-48 h.

Preparation of thermo reversible buffered gel containing PLGA nanoparticles

Poly(ethylene glycol)-block-poly(propylene glycol-blockpoly(ethylene glycol) (20% w/v) was added in 4.2 citrate buffer and stored at 2-8o C temperature for 24 hour for complete dissolution than 2.5% w/v of Pluronic® F-108 (poly(ethylene glycol-block-poly(propylene glycol-block poly(ethylene glycol) added to it and stored further 12 h. Liquid slight viscous solution was obtained. This solution was checked for gel formation by gradually increasing temperature in water bath and observed visually. Dried TNF loaded PLGA nanoparticles were added to this solution and stored in freeze until further use.

Nanoparticle characterization

Particle size determination: The particles size and distribution of TNF loaded PLGA nanoparticles were measured by dynamic light scattering technique using Brookhaven 90plus, USA at 25o C. Particles size was analyzed before lyophilization of the batches and represented as effective diameter in nm.

Morphology: The micro scaled image was taken by scanning electron microscope. The lyophilized powder was sprinkle on carbon tap stick stub than it was coated by gold and focused under microscope under vacuum and 5KV than focused images was captured at different magnification.

Encapsulation efficiency (EE) and drug loading (DL): EE & DL were determined by RP-HPLC method. 10 mg of lyophilized TNF loaded PLGA nanoparticles were dissolved in 2 ml ethyl acetate after dissolving completely, 1 ml HPLC water added and mix well and ultrasonicated for 1 min. for drug extraction and dissolution. Solvent was evaporated stirring overnight at room temperature. The sample filtered by 0.22 µm Durapore (PVDF) membrane filter than injected by auto sampler and TNF concentration was calculated by AUC of peak using standard curve. The %EE & %DL were calculated by using following formulas:

%Drug loading :( Drug in nanoparticles /Weight of nanoparticles) X 100

% Encapsulation efficiency: (Total drug in nanoparticles / Total drug taken) X 100

In vitro drug release from PLGA nanoparticles dispersed in buffered gel: In vitro release of TNF from the PLGA (75:25) nanoparticles was conducted over 24 h using Simulated Vaginal fluid type 1 (SVF). 50 mg of PLGA nanoparticles (159.5 µg TNF) was added to 5 ml thermoreversible buffered gel and dispersed by shaking or using slight vortex than nanoparticles dispersed thermoreversible citrate buffered gel was added to 20 ml SVF as a release medium in a 50 ml tube than placed in shaker (MaxQ4000) at 37 °C with an agitation speed of 60 rpm. At predetermined time intervals, 1 ml of release medium and replaced with fresh medium. The amount of drug released was measured by RP-HPLC method at 260 nm. The samples was filtered by syringe driven Durapore (PVDF) 0.22 um membrane filter before injecting in to HPLC. The standard curve was prepared using mobile phase consisted of acetonitrile/50mM sodium phosphate buffer pH 5.12 at a ratio of (2.5:97.5; v/v). The flow rate was 1 ml/min. and injection volume was 20 µl. Linearity was obtained by 2-10 µg/ml (r2 0.9988) with calibration curve of Area = 49185Conc. + 865.86.

 

RESULTS AND DISCUSSION

Two different methods were applied for preparation of PLGA nanoparticles with TNF. In first method (Solvent diffusion) effective particle diameters of nanoparticles without and with TNF were 94.3-137 nm (PGSD1) and 207.8-228.2 nm (PGSD4) respectively with narrow size distribution (Table 1, Figure 1,2). The drawback of this method was found poor encapsulation of TNF with PLGA nanoparticles, when it was analyzed by RP-HPLC, no drug peak was detected. On the theoretical basis small amount of TNF should be encapsulated during nanoprecipitation, but during our studies we did not observed TNF at quantification level. Possible reason can be partitioning effect towards the aqueous phase during mixing. Another reason may be Triton X 100 that preventing TNF encapsulation because of strong surfactant effect. Therefore, we tried alternative method (emulsion-solvent evaporation) for water soluble drug molecules based on literature [9]. But the particle size was increased from 265 nm to 413 nm as compared to solvent diffusion method and PDI increased from 0.131 to 0.217. Lower particles size was obtained with increased ultra-sonication time [10]. Significant variation in particle size was not observed when Pluronic F68 concentration increased from 0.2 to 1% w/v in aqueous phase. Triton X 100, a non-ionic surfactant was effectively reduced particle size and PDI but TNF encapsulation was not detected in PLGA nanoparticles. Triton X 100 as a surfactant in PLGA nanoparticle preparation was first time tried by our lab and result showed particle size than 100 nm of PLGA nanoparticles that suggesting further use this surfactant with other polymers systems. We did not found any report for use of Triton X 100 in preparation of PLGA nanoparticles.

Encapsulation & drug loading was obtained in range 1.43- 10.78 and 0.25-1.18% respectively. EE was not decreased significantly, when drug concentration increased from 10 mg to 100 mg with 400 mg of PLGA [7]. Double emulsification method is commonly applied technique for encapsulation water soluble drugs in nanoparticles but our results showed low encapsulation of TNF because of the drug rapid partitioning to the external aqueous phase [11, 12].

The thermoreversible gel containing TNF loaded PLGA nanoparticles were developed for vaginal delivery. Poloxamer 407 (20% w/v) & Pluronic® F-108 (2.5% w/v) in combination showed gel matrix formation in between 28-30o C temperature range and reverse into solution form when placed at 2-8o C in refrigerator. Addition of Pluronic®F-108 was increased gelling temperature and viscosity of solution [13,14]. So it can be useful to optimize the gelling temperature close to the body temperature.

SEM image of PLGA nanoparticles showed smooth and spherical in shape (Figure 2). The particles were attached with other because of aggregation during lyophilization. Particles were seen properly when they coated with gold before focusing under electron microscope. The dark spot was observed when high voltage electron beam was applied on PLGA nanoparticles. It may be due to degradation of PLGA when exposed to high energy beam.

In-vitro release of TNF in SVF conducted for 24 h (Figure 3,4). First half an hour release was 15% that may be because of local bust release from the outer gel matrix. In 8 h, cumulative % drug release was 61% and after 24 h it was 67.75%. In-vitro release data is clearly indicating sustained release profile over the 24 h. Gel matrix mass was reduced over the 24 h release study that showed matrix erosion. Drug Release kinetics such as zero order, first order, and Higuchi model were fitted to release profile and R2 value was obtained 0.852, 0.961, and 0.982 respectively. Higuchi model shows that drug released by diffusion mechanism. Korsmeyer-Peppas model fitted to release profile, n-value was 0.45. Peppas model showed Non -Fickian transport that is indicative of the release mechanism from diffusion and erosion controlled process [9].

Table 1: Batch variables and Physiochemical parameters of PLGA nanoparticles.

Batch no. PLGA conc. (mg) TNF (mg) Triton X100 Pluronic F68 (%w/v) Sonication time (min. Particle size (nm) PDI EE (%) DL (%) Zeta Potential (mV)
PGSD1 100 0 0.5 0 0 97 0.074 - - -
PGSD2 400 0 0.5 0 0 137.1 0.062 - - -
PGSD3 400 10 0.5 0 0 207.8 0.110 - - -
PGSD4 100 10 0 0.2 0 228.2 0.143 - - -
PGDE1 100 10 0 0.2 5 413.6 0.237 10.78 1.18 -
PGDE2 400 0 0 0.6 10 278 0.131 - - -5.86
PGDE3 400 100 0 0.6 10 298 0.217 1.18 0.31 -6.53
PGDE4 400 100 0 1 10 300 0.209 1.43 0.42 -5.76

 

CONCLUSION

Both preparation methods were efficient to prepare PLGA nanoparticles. Results revealed that emulsification-solvent evaporation method has advantage over the solvent diffusion method for encapsulation of TNF but particles size was increased as compare to solvent diffusion method. Thermoreversible solution dispersed TNF loaded PLGA nanoparticles was slight viscous at 2-8o C temperature and have capability to converted to gel matrix at 28-30o temperature. More concentration variation in combination of Poloxamer 407 & Pluronic® F-108 can help to optimize gelling temperature close to vagina temperature. Sustained drug release behavior was showed by in-vitro release data and drug release mechanism was diffusion and erosion controlled. This study suggested that possibility of this formulation for vaginal drug delivery of anti HIV drugs. Further studies are required to optimize formulation with enhanced encapsulation, stability and in vivo efficacy in animal models. Additionally, it is required to generate toxicity & safety data.

ACKNOWLEDGEMENTS

This project has been funded in whole or in part with Federal funds (UL1TR000101 previously UL1RR031975) from the National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, through the Clinical and Translational Science Awards Program (CTSA), a trademark of DHHS, part of the Roadmap Initiative, “Re-Engineering the Clinical Research Enterprise.

REFERENCES

1. Hawkins T, Veikley W, St Claire RL 3rd, Guyer B, Clark N, Kearney BP. Intracellular pharmacokinetics of tenofovir diphosphate, carbovir triphosphate, and lamivudine triphosphate in patients receiving triple-nucleoside regimens. J Acquir Immune Defic Syndr. 2005; 39: 406-11.

2. Tenofovir. Leading HIV Medication, Linked with Risk of Kidney Damage. 2012; 415-221.

3. Azanza JR, García Quetglas E, Sádaba B, Gómez-Giu A. Tenofovir: pharmacology and interactions. Enferm Infecc Microbiol Clin. 2008; 26: 2-6.

4. Gitman MD, Hirschwerk D, Baskin CH, Singhal PC. Tenofovir-induced kidney injury. Tenofovir-induced kidney injury. 2007; 6: 155-64.

5. James C, Steinhaus M, Szabo S, Dressler R. Tenofovir-related nephrotoxicity: case report and review of the literature. Pharmacotherapy. 2004; 24: 515-518.

6. Farokhzad OC, Cheng J, Teply BA, Sherifi I, Jon S, Kantoff PW, et al. Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. Proc Natl Acad Sci U S A. 2006; 103: 6315-20.

7. Zhang T, Sturgis TF, Youan BB. pH-responsive nanoparticles releasing tenofovir intended for the prevention of HIV transmission. Eur J Pharm Biopharm. 2011; 79: 526–536.

8. Xu X, Khan MA, Burgess DJ. A quality by design (QbD) case study on liposomes containting hydrophilic API: I formulation, processing design and risk assessment. Int J Pharm. 2011; 419: 52-59.

9. Amjadi I, Rabiee M, Hosseini MS, Mozafari M. Synthesis and characterization of Doxorubicin-loaded Poly(Lactide-co-glycolide) nanoaprticles as a sustained release anticancer drug delivery system. Appl Biotechnol. 2012; 168: 1434-1447.

10. Cohen-Sela E, Chorny M, Koroukhov N, Danenberg HD, Golomb G. A new double emulsion solvent diffusion technique for encapsulating hydrophilic molecules in PLGA nanoparticles. J Control Release. 2009; 133: 90–95.

11. Tewes F, Munnier E, Antoon B, Ngaboni Okassa L, Cohen-Jonathan S, Marchais H, et al. Comparative study of doxorubicin-loaded poly(lactide-co-glycolide) nanoparticles prepared by single and double emulsion methods. Eur J Pharm Biopharm. 2007; 66: 488-92.

12. Azizi M, Farahmandghavi F, Joghataei M, Zandi M, Imani M, Bakhtiary M, et al. Fabrication of protein-loaded PLGA nanoparticles: effect of selected formulation variables on particle size and release profile. J Polym Res. 2013; 20:110-124.

13. Aka-Any-Grah A, Bouchemal K, Koffi A, Agnely F, Zhang M, Djabourov M, Ponchel G. Formulation of mucoadhesive vaginal hydrogels insensitive to dilution with vaginal fluids. Eur J Pharm Biopharm. 2010; 76: 296–303.

14. Date AA, Shibata A, Goede M, Sanford B, La Bruzzo K, Belshan M, et al. Development and evaluation of a thermosensitive vaginal gel containing raltegravir + efavirenz loaded nanoparticles for HIV prophylaxis. Antiviral Res. 2012; 96: 430–436.

Abstract

This study was designed to develop buffered thermoreversible gel containing tenofovir (TNF) loaded PLGA nanoparticles for sustained drug release and prolonged protection from HIV-1 sexual transmission. Optimized buffered thermoreversible formulation containing TNF loaded PLGA nanoparticles was in solution form at 2-8oC and converted into a gel matrix at 28oC. PLGA nanoparticles were prepared by 
two methods; (a) Solvent diffusion method, (b) Emulsification- solvent evaporation and effective particles diameter was 94.3-228 nm and 265-413 nm respectively by dynamic light scattering technique. Morphology of the particles was analyzed by scanning electron microscope and micro image showed smooth & spherical in shape nanoparticles. Triton® X-100 a non-ionic surfactant effectively reduced the particle size of nanoparticles in solvent diffusion method but the desired TNF encapsulation was not observed. Drug encapsulation and drug loading were analyzed in nanoparticles prepared by emulsification-solvent evaporation and the values were 1.43-10.78% and 0.25-1.18% respectively. In-vitro release of TNF from Buffered thermoreversible gel containing PLGA nanoparticles was conducted in simulated vaginal fluid (pH 4.2). In 8 h, cumulative % drug release was 61% and after 24 h it was 67.75%. Drug release mechanism was diffusion and erosion controlled. Results demonstrated the therapeutic potential of the vaginal nano gel formulation for prolonged protection against HIV-1 transmission as an STD.

Citation

Karla PK (2013) Thermo Reversible Buffered Tenofovir Nano Gel for Vaginal Delivery. J Pharmacol Clin Toxicol 1(2): 1010.

Received : 02 Sep 2013
Accepted : 04 Nov 2013
Published : 06 Nov 2013
Journals
Annals of Otolaryngology and Rhinology
ISSN : 2379-948X
Launched : 2014
JSM Schizophrenia
Launched : 2016
Journal of Nausea
Launched : 2020
JSM Internal Medicine
Launched : 2016
JSM Hepatitis
Launched : 2016
JSM Oro Facial Surgeries
ISSN : 2578-3211
Launched : 2016
Journal of Human Nutrition and Food Science
ISSN : 2333-6706
Launched : 2013
JSM Regenerative Medicine and Bioengineering
ISSN : 2379-0490
Launched : 2013
JSM Spine
ISSN : 2578-3181
Launched : 2016
Archives of Palliative Care
ISSN : 2573-1165
Launched : 2016
JSM Nutritional Disorders
ISSN : 2578-3203
Launched : 2017
Annals of Neurodegenerative Disorders
ISSN : 2476-2032
Launched : 2016
Journal of Fever
ISSN : 2641-7782
Launched : 2017
JSM Bone Marrow Research
ISSN : 2578-3351
Launched : 2016
JSM Mathematics and Statistics
ISSN : 2578-3173
Launched : 2014
Journal of Autoimmunity and Research
ISSN : 2573-1173
Launched : 2014
JSM Arthritis
ISSN : 2475-9155
Launched : 2016
JSM Head and Neck Cancer-Cases and Reviews
ISSN : 2573-1610
Launched : 2016
JSM General Surgery Cases and Images
ISSN : 2573-1564
Launched : 2016
JSM Anatomy and Physiology
ISSN : 2573-1262
Launched : 2016
JSM Dental Surgery
ISSN : 2573-1548
Launched : 2016
Annals of Emergency Surgery
ISSN : 2573-1017
Launched : 2016
Annals of Mens Health and Wellness
ISSN : 2641-7707
Launched : 2017
Journal of Preventive Medicine and Health Care
ISSN : 2576-0084
Launched : 2018
Journal of Chronic Diseases and Management
ISSN : 2573-1300
Launched : 2016
Annals of Vaccines and Immunization
ISSN : 2378-9379
Launched : 2014
JSM Heart Surgery Cases and Images
ISSN : 2578-3157
Launched : 2016
Annals of Reproductive Medicine and Treatment
ISSN : 2573-1092
Launched : 2016
JSM Brain Science
ISSN : 2573-1289
Launched : 2016
JSM Biomarkers
ISSN : 2578-3815
Launched : 2014
JSM Biology
ISSN : 2475-9392
Launched : 2016
Archives of Stem Cell and Research
ISSN : 2578-3580
Launched : 2014
Annals of Clinical and Medical Microbiology
ISSN : 2578-3629
Launched : 2014
JSM Pediatric Surgery
ISSN : 2578-3149
Launched : 2017
Journal of Memory Disorder and Rehabilitation
ISSN : 2578-319X
Launched : 2016
JSM Tropical Medicine and Research
ISSN : 2578-3165
Launched : 2016
JSM Head and Face Medicine
ISSN : 2578-3793
Launched : 2016
JSM Cardiothoracic Surgery
ISSN : 2573-1297
Launched : 2016
JSM Bone and Joint Diseases
ISSN : 2578-3351
Launched : 2017
JSM Bioavailability and Bioequivalence
ISSN : 2641-7812
Launched : 2017
JSM Atherosclerosis
ISSN : 2573-1270
Launched : 2016
Journal of Genitourinary Disorders
ISSN : 2641-7790
Launched : 2017
Journal of Fractures and Sprains
ISSN : 2578-3831
Launched : 2016
Journal of Autism and Epilepsy
ISSN : 2641-7774
Launched : 2016
Annals of Marine Biology and Research
ISSN : 2573-105X
Launched : 2014
JSM Health Education & Primary Health Care
ISSN : 2578-3777
Launched : 2016
JSM Communication Disorders
ISSN : 2578-3807
Launched : 2016
Annals of Musculoskeletal Disorders
ISSN : 2578-3599
Launched : 2016
Annals of Virology and Research
ISSN : 2573-1122
Launched : 2014
JSM Renal Medicine
ISSN : 2573-1637
Launched : 2016
Journal of Muscle Health
ISSN : 2578-3823
Launched : 2016
JSM Genetics and Genomics
ISSN : 2334-1823
Launched : 2013
JSM Anxiety and Depression
ISSN : 2475-9139
Launched : 2016
Clinical Journal of Heart Diseases
ISSN : 2641-7766
Launched : 2016
Annals of Medicinal Chemistry and Research
ISSN : 2378-9336
Launched : 2014
JSM Pain and Management
ISSN : 2578-3378
Launched : 2016
JSM Women's Health
ISSN : 2578-3696
Launched : 2016
Clinical Research in HIV or AIDS
ISSN : 2374-0094
Launched : 2013
Journal of Endocrinology, Diabetes and Obesity
ISSN : 2333-6692
Launched : 2013
Journal of Substance Abuse and Alcoholism
ISSN : 2373-9363
Launched : 2013
JSM Neurosurgery and Spine
ISSN : 2373-9479
Launched : 2013
Journal of Liver and Clinical Research
ISSN : 2379-0830
Launched : 2014
Journal of Drug Design and Research
ISSN : 2379-089X
Launched : 2014
JSM Clinical Oncology and Research
ISSN : 2373-938X
Launched : 2013
JSM Bioinformatics, Genomics and Proteomics
ISSN : 2576-1102
Launched : 2014
JSM Chemistry
ISSN : 2334-1831
Launched : 2013
Journal of Trauma and Care
ISSN : 2573-1246
Launched : 2014
JSM Surgical Oncology and Research
ISSN : 2578-3688
Launched : 2016
Annals of Food Processing and Preservation
ISSN : 2573-1033
Launched : 2016
Journal of Radiology and Radiation Therapy
ISSN : 2333-7095
Launched : 2013
JSM Physical Medicine and Rehabilitation
ISSN : 2578-3572
Launched : 2016
Annals of Clinical Pathology
ISSN : 2373-9282
Launched : 2013
Annals of Cardiovascular Diseases
ISSN : 2641-7731
Launched : 2016
Journal of Behavior
ISSN : 2576-0076
Launched : 2016
Annals of Clinical and Experimental Metabolism
ISSN : 2572-2492
Launched : 2016
Clinical Research in Infectious Diseases
ISSN : 2379-0636
Launched : 2013
JSM Microbiology
ISSN : 2333-6455
Launched : 2013
Journal of Urology and Research
ISSN : 2379-951X
Launched : 2014
Journal of Family Medicine and Community Health
ISSN : 2379-0547
Launched : 2013
Annals of Pregnancy and Care
ISSN : 2578-336X
Launched : 2017
JSM Cell and Developmental Biology
ISSN : 2379-061X
Launched : 2013
Annals of Aquaculture and Research
ISSN : 2379-0881
Launched : 2014
Clinical Research in Pulmonology
ISSN : 2333-6625
Launched : 2013
Journal of Immunology and Clinical Research
ISSN : 2333-6714
Launched : 2013
Annals of Forensic Research and Analysis
ISSN : 2378-9476
Launched : 2014
JSM Biochemistry and Molecular Biology
ISSN : 2333-7109
Launched : 2013
Annals of Breast Cancer Research
ISSN : 2641-7685
Launched : 2016
Annals of Gerontology and Geriatric Research
ISSN : 2378-9409
Launched : 2014
Journal of Sleep Medicine and Disorders
ISSN : 2379-0822
Launched : 2014
JSM Burns and Trauma
ISSN : 2475-9406
Launched : 2016
Chemical Engineering and Process Techniques
ISSN : 2333-6633
Launched : 2013
Annals of Clinical Cytology and Pathology
ISSN : 2475-9430
Launched : 2014
JSM Allergy and Asthma
ISSN : 2573-1254
Launched : 2016
Journal of Neurological Disorders and Stroke
ISSN : 2334-2307
Launched : 2013
Annals of Sports Medicine and Research
ISSN : 2379-0571
Launched : 2014
JSM Sexual Medicine
ISSN : 2578-3718
Launched : 2016
Annals of Vascular Medicine and Research
ISSN : 2378-9344
Launched : 2014
JSM Biotechnology and Biomedical Engineering
ISSN : 2333-7117
Launched : 2013
Journal of Hematology and Transfusion
ISSN : 2333-6684
Launched : 2013
JSM Environmental Science and Ecology
ISSN : 2333-7141
Launched : 2013
Journal of Cardiology and Clinical Research
ISSN : 2333-6676
Launched : 2013
JSM Nanotechnology and Nanomedicine
ISSN : 2334-1815
Launched : 2013
Journal of Ear, Nose and Throat Disorders
ISSN : 2475-9473
Launched : 2016
JSM Ophthalmology
ISSN : 2333-6447
Launched : 2013
Annals of Psychiatry and Mental Health
ISSN : 2374-0124
Launched : 2013
Medical Journal of Obstetrics and Gynecology
ISSN : 2333-6439
Launched : 2013
Annals of Pediatrics and Child Health
ISSN : 2373-9312
Launched : 2013
JSM Clinical Pharmaceutics
ISSN : 2379-9498
Launched : 2014
JSM Foot and Ankle
ISSN : 2475-9112
Launched : 2016
JSM Alzheimer's Disease and Related Dementia
ISSN : 2378-9565
Launched : 2014
Journal of Addiction Medicine and Therapy
ISSN : 2333-665X
Launched : 2013
Journal of Veterinary Medicine and Research
ISSN : 2378-931X
Launched : 2013
Annals of Public Health and Research
ISSN : 2378-9328
Launched : 2014
Annals of Orthopedics and Rheumatology
ISSN : 2373-9290
Launched : 2013
Journal of Clinical Nephrology and Research
ISSN : 2379-0652
Launched : 2014
Annals of Community Medicine and Practice
ISSN : 2475-9465
Launched : 2014
Annals of Biometrics and Biostatistics
ISSN : 2374-0116
Launched : 2013
JSM Clinical Case Reports
ISSN : 2373-9819
Launched : 2013
Journal of Cancer Biology and Research
ISSN : 2373-9436
Launched : 2013
Journal of Surgery and Transplantation Science
ISSN : 2379-0911
Launched : 2013
Journal of Dermatology and Clinical Research
ISSN : 2373-9371
Launched : 2013
JSM Gastroenterology and Hepatology
ISSN : 2373-9487
Launched : 2013
Annals of Nursing and Practice
ISSN : 2379-9501
Launched : 2014
JSM Dentistry
ISSN : 2333-7133
Launched : 2013
Author Information X