Protein malnutrition in BALB/C mice: A model mimicking clinical scenario of marasmic-kwashiorkor malnutrition.

Protein malnutrition continues to be a major global issue. A stable animal model to address protein malnutrition and its effect on various disease conditions is necessary. In the present study, we have formulated and standardized a low protein diet (LPD) to develop a protein malnutrition model using Balb/C mice. Healthy male Balb/C mice were weaned and exposed to LPD combinations while another group exposed to normal diet (18% protein). Animal survival, change in body weight, body mass index (BMI), biochemical parameters, antioxidant status, and liver histopathology were used to confirm the development of malnourished mice model (marasmic-kwashiorkor). Mice receiving 10% protein diet showed moderate weight gain, higher BMI, and no mortality compared to the 6% protein group. The former group showed remarkable differences in BMI, biochemical and antioxidant parameters. Further, histopathological changes against the normal group at weeks 20 and 30 confirmed the development of protein malnutrition in mice on 10% protein diet. The study confirms the development of a stable, economical, reproducible, and clinically relevant protein malnutrition model using the formulated 10% protein diet. Further, the model can be used for short and long-term studies to investigate the pathophysiology of malnutrition in any disease/condition.

Drug Release and Cytotoxicity of Hyaluronic Acid and Zinc Oxide Gels, An In-Vitro Study.

Hyaluronic acid (HA) is a naturally occurring biopolymer, with a remarkable wound healing property. Zinc-oxide non-eugenol is a material widely used for periodontal dressing in dentistry. However, it has been reported that zinc oxide non-eugenol is toxic to osteoblasts and fibroblasts. Hence, the present study aimed to evaluate the drug release and cytotoxicity of HA and zinc-oxide gels. Hydrogels of HA and zinc oxide were formulated with carbopol as a carrier. In vitro drug release was performed by UV spectrophotometry, dialysis, and vial bag methods. Cytotoxicity assessment of HA and zinc-oxide gels was performed in human periodontal ligament fibroblasts (HPdLF) and human gingival fibroblasts (hGFs). An inverted phase-contrast microscope was used to assess the morphological changes. At 24 and 48 hr, HPdLF cells showed the highest viability in 0.1% low molecular weight-HA (LMW-HA) with a median value of 131.9, and hGFs showed the highest viability in 5% LMW-HA with a median of 129.56. The highest viability of HPdLF cells was observed in 5% high molecular weight-HA (HMW-HA), with a median value of 127.11. hGFs showed the highest viability in 1% HMW-HA with a median value of 97.99. Within the limitations of the present study, we concluded that LMW-HA is more efficient than HMW-HA. Both HPdLF and hGF cells showed complete cell morbidity with zinc-oxide hydrogels. Therefore, zinc oxide-based gels in concentrations as low as 9% could be toxic intraorally to soft tissues that harbor gingival and periodontal ligament fibroblasts.

Influence of in vitro oxygen concentrations on preimplantation embryo development, gene expression and production of Hanwoo calves following embryo transfer.

This study evaluated the effects of two different oxygen (O2) concentrations on in vitro embryo development, embryo quality, and gene expression and the in vivo development following embryos transfer to recipients of natural and synchronized estrus in bovines. Cumulus oocyte complexes were in vitro matured in TCM199 supplemented with FSH (10 microg/ml), LH (10 microg/ml), and 10% (v/v) FBS. Presumptive zygotes were cultured in SOF medium either under 5% (low) or 20% (high) O2 in air. Cleavage rates did not differ between groups. Blastocyst and hatched blastocyst development in 5% O2 were significantly (P < 0.05) higher than in 20% O2. Total cell number of in vivo blastocyst was significantly (P < 0.05) higher than that of in vitro blastocyst. ICM ratio and apoptosis of in vivo blastocyst were significantly (P < 0.05) lower than that of in vitro blastocyst. Using real time PCR, we have found that for the set of genes (GLUT-1, MnSOD, VEGF, Bax, and Bcl-2) analyzed, there were differences in mRNA expression between in vitro produced (IVP) and in vivo produced embryos. Interestingly, the abundance of transcript for IFN-tau in IVP embryos produced under 5% O2 concentration was similar to in vivo counterparts. The pregnancy and twin rates of natural recipients were significantly (P < 0.05) higher than those of synchronized counterparts. No significant difference in the offspring sex was observed. In conclusion, low (5%) O2 concentration during IVC was beneficial for enhancing the embryo quality and recipients of natural estrus were more suitable than synchronized estrus for stable production of Hanwoo calves.