07272nam a22001937a 4500 291826 291826 633.8 APA/PR Ph.D Aparna, P M Propagation and bioactivity studies in Ellotti (Pterospermum rubiginosum b. Heyne ex wight & arn.) Vellanikkara Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture 2024 173,xiip. Ph.D The study entitled “Propagation and bioactivity studies in Ellotti (Pterospermum rubiginosum B. Heyne ex Wight & Arn.)” was conducted at the Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture, KAU, Vellanikkara, and the College of Veterinary and Animal Sciences, KVASU, Pookode, Wayanad from 2018 to 2023. The study aimed to collect ethnobotanical information from tribal healers in Kerala, develop vegetative propagation protocols, and investigate the bioactivity of Pterospermum rubiginosum through antioxidant, antimicrobial, wound healing, and antihyperglycemic studies. From the account of tribal healers, the bark of Pterospermum rubiginosum is an effective remedy for the treatment of fractures. The tree's bark is effectively used by the ethnomedical practitioners of different tribes in Kerala like Kurichiyan, Kattunayakan, Muthuvan, Vetta Kuruman and Kanikkar. These experienced healers assert that majority of patients respond well to the treatment, approximately one in a hundred might exhibit allergic reactions or fail to respond altogether. It may take a week to three months for a broken bone to mend. For a fracture to heal, all factors like time taken for the patient to reach the healer, severity of the fracture, patient's age, calcium status in the body and other health conditions should be considered. In the vegetative propagation study conducted during June-July and September-October, maximum rooting was observed in the June-July months, with the highest rooting percentage of 4.53 per cent. Among the growth regulators, NAA 1000 ppm was found to be significantly superior in enchaining the rooting in both hardwood and semi-hardwood cuttings of Pterospermum rubiginosum at 120 days after planting. Biochemical analysis of bark and leaves of Pterospermum rubiginosum revealed the presence of phenols, flavonoids, tannins, steroids, saponins, sugars and carbohydrates with the bark showing higher values compared to the leaves. The gas chromatography-high resolution mass spectrometry (GC-HRMS) and high resolution -liquid chromatography mass spectrometry (HR-LCMS) analysis of ethanol extracts of bark and leaves revealed the presence of 22 major compounds in the bark extract and 32 major compounds in the leaf extract. The majority of the abundant compounds belong to flavonoid, phenol, alkaloid and saponin class of compounds like 6''-caffeoylisoorientin, kaempferol 3-O-β-Dgalactoside, 3-O-cis-coumaroylmaslinic acid, epigallocatechin and patientoside A. The bark extract exhibited higher antioxidant activity in 2,2-diphenyl-2- picrylhydrazyl hydrate (DPPH) radical scavenging assay, 2,2–azino–bis (3- ethylbenzothiozoline-6-sulfonic acid) radical scavenging assay (ABTS), reducing power assay, ferric reducing antioxidant power (FRAP) assay and total antioxidant activity compared to the leaf extract, and similar activity to that of leaf extract in nitric oxide scavenging assay. The ethanol extract of both bark and leaves of Pterospermum rubiginosum were ineffective against Staphylococcus aureus and Escherichia coli up to a concentration of 1000 μg/mL in Kirby-Bauer agar disc diffusion method. However, in modified resazurin microtiter plate assay, the ethanol extract of Pterospermum rubiginosum bark exhibited minimum inhibitory concentration of 500 μg/mL against Staphylococcus aureus, while the leaf extract showed inhibition against Escherichia coli at a minimum inhibitory concentration of 1750 μg/mL. The ethanol extract of Pterospermum rubiginosum bark was used for further in vitro studies as it showed superior biochemical, antioxidant and antimicrobial properties compared to leaf extract. The per cent mean cell viability of ethanol extract of Pterospermum rubiginosum bark in L929 mouse skin fibroblast cells was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and resulted an effective concentration 50 (EC50) of 100.90 μg/mL. In scratch assay using L929 mouse skin fibroblast cells, gap closure was at a faster rate in positive control, followed by test drug and normal control. The in vitro test was followed by an in vivo wound healing study in Wistar rats. The per cent reduction in wound area on the 15th day of wound induction in Wistar rats was similar in Cipladine standard (89.77 ± 2.39) and test extracts group at 2.5 per cent (86.99 ± 1.17), 5.0 per cent (87.25 ± 1.91) and 10.0 per cent (85.99 ± 1.01), which were significantly higher than normal control group (78.54 ± 2.93). The presence of keratinocytes and incompletely formed hair follicles in histopathological observations of Cipladine standard and extract 2.5 per cent group suggested that the animals were in proliferative phase of wound healing whereas, incomplete epithelialization in extracts 5.0 per cent, 10.0 per cent and normal control led to the conclusion that the animals were in inflammatory phase of wound healing. In Wistar rats, further analysis using the oral glucose tolerance test (OGTT) revealed that glucose values were higher one hour after glucose administration and followed by a steady decrease after one hour. After three hours, the lowest glucose values were observed in glibenclamide standard and extract at 50 mg/kg body weight groups, whereas, normal control and higher concentration groups exhibited significantly higher glucose values. The study concludes that tribal healers effectively utilise Pterospermum rubiginosum bark for healing bone fractures, wound and sprains. The ethanol extract of Pterospermum rubiginosum bark and leaves exhibited antioxidant and antimicrobial activity. In vivo wound healing studies suggested that a lower concentration of extract at 2.5 per cent was effective in wound healing compared to higher concentrations, possibly due to the presence of compounds hindering the migration of proinflammatory cytokines to the wound site. Complete wound healing could have been observed if the duration of study was increased to a few more days. Another significant finding was the reduction in glucose level in Wistar rats after administration of extract at 50 mg/kg body weight, which indicated that Pterospermum rubiginosum has anti-hyperglycemic properties. Further in vivo wound healing studies could be carried out by using bark extract at lower concentrations or by isolation of compounds responsible for wound healing. Plantation Spices Medicinal and Aromatic crops Ellotti Pterospermum rubiginosum b. Heyne ex wight & arn. Vikram, H C(Guide) https://krishikosh.egranth.ac.in/handle/1/5810219827 ddc TH 0 0 0 1 TH KAUCLV KAUCLV THESES 2025-01-21 0 633.8 APA/PR Ph.D 176235 2025-01-21 00:00:00 TH