Fabrication of ethosuximide loaded alginate/polyethylene oxide scaffolds for epilepsy research using 3D-printing method.

Epilepsy is a medical condition that causes seizures and impairs the mental and physical activities of patients. Unfortunately, over one-third of patients do not receive adequate relief from oral Antiepileptic Drugs (AEDs) and continue to experience seizures. In addition to that, long term usage of Antiepileptic Drugs can cause a range of side effects. To overcome this problem, the precision of 3D printing technology is combined with the controlled release capabilities of biodegradable polymers, allowing for tailored and localized AED delivery to specific seizure sites. As a result of this novel technique, therapeutic outcomes can be enhanced, side effects of AEDs are minimized, and patient-specific dosage forms can be created. This study focused on the use of ethosuximide, an antiepileptic drug, at different concentrations (10, 13, and 15 mg) loaded into 3D-printed sodium alginate and polyethylene oxide scaffolds. The scaffolds contained varying concentrations (0.25%, 0.50%, and 0.75% w/v) and had varying pores created by 3D patterning sizes from 159.86 ± 19.9 µm to 240.29 ± 10.7 µm to optimize the releasing system for an intracranial administration. The addition of PEO changed the Tg and Tm temperatures from 65°C to 69°C and from 262°C to 267°C, respectively. Cytotoxicity assays using the human neuroblastoma cell line (SH-SY5Y) showed that cell metabolic activity reached 130% after 168 h, allowing the cells to develop into mature neural cells. In vitro testing demonstrated sustained ethosuximide release lasting 2 hours despite crosslinking with 3% CaCl2. The workpaves the way for the use of ethosuximide -loaded scaffolds for treating epilepsy.

Biodegradable Multi-layered Silk Fibroin-PCL Stent for the Management of Cervical Atresia: In Vitro Cytocompatibility and Extracellular Matrix Remodeling In Vivo.

Cervical atresia is a rare congenital Müllerian duct anomaly that manifests as the absence or deformed nonfunctional presence of the cervix. Herein, a multi-layered biodegradable stent is fabricated using a homogeneous blend of silk fibroin with polycaprolactone using hexafluoroisopropanol as a common solution. Briefly, a concentric cylinder of 3D honeycomb layer is sandwiched within electrospun sheets for fixing at the cervico-uterine junction to pave the way of cervical reconstruction. An average length of 40 mm with 3 mm diameter is fabricated for the hybrid stent design. SEM evidences an evenly distributed pore architecture of the electrospun layer, and mechanical characterization of stent reveals a tensile strength of 1.7 ± 0.2 MPa, with a Young's modulus of 5.9 ± 0.1 MPa. Physico-chemical characterization confirms the presence of silk fibroin and poly caprolactone within the engineered stent. Following 14 days of pepsin enzymatic degradation, 18% degradation and a contact angle measurement of 97° are observed. In vitro cytocompatibility studies are performed using site-specific primary human cervical squamous, columnar epithelial cells, and human endometrial stromal cells. The study demonstrates non-cytotoxic cells' viability (no significant toxicity), improved cell anchoring, adherence among the stent layers, and proliferation in the 3D microenvironment. Furthermore, in vivo subcutaneous studies in the rodent model indicate that the implanted stent undergoes constructive remodeling, neo-tissue creation, neo-vasculature formation, and re-epithelialization while maintaining patency for 2 months.

Accuracy of 3D printed spine models for pre-surgical planning of complex adolescent idiopathic scoliosis (AIS) in spinal surgeries: a case series.

Adolescent idiopathic scoliosis (AIS) is a noticeable spinal deformity in both adult and adolescent population. In majority of the cases, the gold standard of treatment is surgical intervention. Technological advancements in medical imaging and 3D printing have revolutionised the surgical planning and intraoperative decision making for surgeons in spinal surgery. However, its applicability for planning complex spinal surgeries is poorly documented with human subjects. The objective of this study is to evaluate the accuracy of 3D printed models for complex spinal deformities based on Cobb angles between 40° to 95°.This is a retrospective cohort study where, five CT scans of the patients with AIS were segmented and 3D printed for evaluating the accuracy. Consideration was given to the Inter-patient and acquisition apparatus variability of the CT-scan dataset to understand the effect on trueness and accuracy of the developed CAD models. The developed anatomical models were re-scanned for analysing quantitative surface deviation to assess the accuracy of 3D printed spinal models. Results show that the average of the root mean square error (RMSE) between the 3DP models and virtual models developed using CT scan of mean surface deviations for the five 3d printed models was found to be 0.5±0.07 mm. Based on the RMSE, it can be concluded that 3D printing based workflow is accurate enough to be used for presurgical planning for complex adolescent spinal deformities. Image acquisition and post processing parameters, type of 3D printing technology plays key role in acquiring required accuracy for surgical applications.

Design modification of surgical drill bit for final osteotomy site preparation towards improved bone-implant contact.

Implant stability significantly impacts accelerated osseointegration, leading to faster patient recovery. Both primary and secondary stability necessitates superior bone-implant contact influenced by the surgical tool required to prepare the final osteotomy site. Besides, excessive shearing and frictional forces generate heat causing local tissue necrosis. Hence, surgical procedure necessitates proper irrigation with water to minimize heat generation. Notably, the water irrigation system removes bone chips and osseous coagulums, which may help accelerate osseointegration and improve bone-implant contact. The inferior bone-implant contact and thermal necrosis at the osteotomy site are primarily responsible for poor osseointegration and eventual failure. Therefore, optimizing tool geometry is key to minimizing shear force, heat generation, and necrosis during final osteotomy site preparation. The present study explores modified drilling tool geometry, especially cutting edge for osteotomy site preparation. The mathematical modeling is used to find out ideal cutting-edge geometry that facilitates drilling under relatively less operational force (0.55-5.24 N) and torque (98.8-154.5 N-mm) with a significant reduction (28.78%-30.87%) in heat generation. Twenty-three conceivable designs were obtained using the mathematical model; however, only three have shown promising results in static structural FEM platforms. These drill bits are designed for the final drilling operation and need to be carried out during the final osteotomy site preparation.

Biomechanical influence of plate configurations on mandible subcondylar fracture fixation: a finite element study.

Mandible subcondylar fractures have very high complication rate, yet there is no consensus on suitable plate design for optimal patient outcomes. Our study is aimed at comparing single mini, trapezoid, lambda, strut and double mini plates. A finite-element (FE) model of intact mandible was developed based on healthy CT-scan data, which was further virtually osteotomized and fixated with plates. The cortical and cancellous bones were assigned region-specific orthotropic and heterogenous isotropic material properties respectively. The models were subjected to six load cases representing the mastication cycle. Under opposite lateralities, the tensile and compressive mandibular strain distributions were found as the opposite, with tensile strains at the posterior border under ipsilateral molar clenching (RMOL) resulting in lesser mandibular strain in reconstructed mandible with single mini plate under RMOL but highest mandibular strain under the contralateral molar clenching (LMOL). Owing to the reduced mandibular strains under LMOL than RMOL, the contralateral chewing is preferred during the immediate post-surgery period for patients. Under LMOL, the peak von Mises stresses in the plate decreased with increase in the number of screws. Furthermore, the presence of two arms in double mini and trapezoid plates seems beneficial to neutralise the tensile and compressive strains across load cases.

Comparative evaluation of a patient-specific customised plate designs and screws for partial mandibular reconstruction.

Mandibles with odontogenic tumors are often partially reconstructed with a metallic bone graft analogue with dental roots, crowns, along with a customized plate fixed with monocortical or bicortical screws, following resection of the tumor. In this study, two different designs of patient specific customized Ti reconstruction plates, solid and plate with holes, were considered. Fixation through both bicortical and monocortical screw types were investigated. FE models of the reconstructed mandibles were developed to analyse the influence of the plate-screw type combination on the load transfer across the mandibles under a mastication cycle. The effective homogenized orthotropic material properties of the lattice structures with 0.6 mm fibre diameter with 0.5 mm inter-fibre space were assigned to material properties for the bone graft analogue. The study shows that the combination of plate and screw types influences the state of stresses in the reconstructed mandible. Based on the results of this patient specific study, following resection of the tumor, either solid Ti plate with bicortical screws or Ti plate with holes along with monocortical screws may be used for partial mandibulectomy. It should also be noted that stresses in none of the plates or screws exceeded the yield limit for Ti under the mastication cycle indicating that the components are safe for mandibular reconstruction. However, the choice of this combination of reconstruction plates and screws is dependant on the condition and severity of the tumor in the diseased mandible.

Production of 3D Printed Bi-Layer and Tri-Layer Sandwich Scaffolds with Polycaprolactone and Poly (vinyl alcohol)-Metformin towards Diabetic Wound Healing.

Type 2 diabetes mellitus (T2DM) is a chronic disease characterized by impaired insulin secretion, sensitivity, and hyperglycemia. Diabetic wounds are one of the significant complications of T2DM owing to its difficulty in normal healing, resulting in chronic wounds. In the present work, PCL/PVA, PCL/PVA/PCL, and metformin-loaded, PCL/PVA-Met and PCL/PVA-Met/PCL hybrid scaffolds with different designs were fabricated using 3D printing. The porosity and morphological analysis of 3D-printed scaffolds were performed using scanning electron microscopy (SEM). The scaffolds' average pore sizes were between 63.6 ± 4.0 and 112.9 ± 3.0 µm. Molecular and chemical interactions between polymers and the drug were investigated with Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). Mechanical, thermal, and degradation analysis of the scaffolds were undertaken to investigate the physico-chemical characteristics of the scaffolds. Owing to the structure, PCL/PVA/PCL sandwich scaffolds had lower degradation rates than the bi-layer scaffolds. The drug release of the metformin-loaded scaffolds was evaluated with UV spectrometry, and the biocompatibility of the scaffolds on fibroblast cells was determined by cell culture analysis. The drug release in the PCL/PVA-Met scaffold was sustained till six days, whereas in the PCL/PVA-Met/PCL, it continued for 31 days. In the study of drug release kinetics, PCL/PVA-Met and PCL/PVA-Met/PCL scaffolds showed the highest correlation coefficients (R2) values for the first-order release model at 0.8735 and 0.889, respectively. Since the layered structures in the literature are mainly obtained with the electrospun fiber structures, these biocompatible sandwich scaffolds, produced for the first time with 3D-printing technology, may offer an alternative to existing drug delivery systems and may be a promising candidate for enhancing diabetic wound healing.

High fibroin-loaded silk-PCL electrospun fiber with core-shell morphology promotes epithelialization with accelerated wound healing.

Silk fibroin (SF) is a widely explored biopolymer for wound-healing applications due to the presence of amino acids in the biodegradable polymer chain with superior mechanical properties. Herein, a high SF-loaded fibrous matrix along with poly(ε-caprolactone) (PCL) was fabricated using electrospinning of emulsion and blend compositions to modulate nanostructure morphology. A comparative study of the physicomechanical properties of electrospun fibers with emulsion (eS7P3) and homogenous blend (bS7P3) was performed as well. In both compositions, SF loading of up to 70% was successfully achieved in the spun fibers while emulsion yielded core-shell morphology, and the blend resulted in monolith fiber architecture as evidenced by TEM microscopy. Further characterization revealed superior mechanical properties in S7P3 fiber with core-shell morphology, as compared to those in the monolith in terms of a higher degree of crystallinity with Young's modulus of 60 MPa under tensile test and nanoindentation modulus of 1.59 ± 0.8 GPa. Further, eS7P3 nanostructure morphology containing silk in the core with a thin outer layer of PCL facilitated relatively faster biodegradation in the lysozyme medium, as compared to that in the monolith. Owing to the presence of a hydrophobic shell, protein adsorption on the fibrous mat presented slow but steady kinetics up to 24 h. When the scaffold was seeded with human placenta-derived mesenchymal stem cells (hPMSCs), in vitro study confirmed that the eS7P3 structure had marginally higher cell proliferation with superior cell infiltration than the monolith. Further, in vivo study involving a rodent model showed the potential of the eS7P3 fiber substrate with a core-shell structure for accelerating full-thickness wound healing by inducing hair follicle and wound closure with less scar formation after 15 days.

Metal Ion Augmented Mussel Inspired Polydopamine Immobilized 3D Printed Osteoconductive Scaffolds for Accelerated Bone Tissue Regeneration.

Critical bone defects with a sluggish rate of auto-osteoconduction and imperfect reconstruction are motivators for the development of an alternate innovative approach for the regeneration of bone. Tissue engineering for bone regeneration signifies an advanced way to overcome this problem by creating an additional bone tissue substitute. Among different fabrication techniques, the 3D printing technique is obviously the most efficient and advanced way to fabricate an osteoconductive scaffold with a controlled porous structure. In the current article, the polycarbonate and polyester diol based polyurethane-urea (P12) was synthesized and 3D porous nanohybrid scaffolds (P12/TP-nHA) were fabricated using the 3D printing technique by incorporating the osteoconductive nanomaterial titanium phosphate adorned nanohydroxyapatite (TP-nHA). To improve the bioactivity, the surface of the fabricated scaffolds was modified with the immobilized biomolecule polydopamine (PDA) at room temperature. XPS study as well as the measurement of surface wettability confirmed the higher amount of PDA immobilization on TP-nHA incorporated nanohybrid scaffolds through the dative bone formation between the vacant d orbital of the incorporated titanium ion and the lone pair electron of the catechol group of dopamine. The incorporated titanium phosphate (TP) increased the tensile strength (53.1%) and elongation at break (96.8%) of the nanohybrid composite as compared to pristine P12. Moreover, the TP incorporated nanohybrid scaffold with calcium and phosphate moieties and a higher amount of immobilized active biomolecule improved the in vitro bioactivity, including the cell viability, cell proliferation, and osteogenic gene expression using hMSCs, of the fabricated nanohybrid scaffolds. A rat tibia defect model depicted that the TP incorporated nanohybrid scaffold with immobilized PDA enhanced the in vivo bone regeneration ability compared to the control sample without revealing any organ toxicity signifying the superior osteogenic bioactivity. Thus, a TP augmented polydopamine immobilized polyurethane-urea based nanohybrid 3D printed scaffold with improved physicochemical properties and osteogenic bioactivity could be utilized as an excellent advanced material for bone regeneration substitute.

Three in One: Stimuli-Responsive Fluorescence, Solid-State Emission, and Dual-Organelle Imaging Using a Pyrene-Benzophenone Derivative.

Small organic luminogens, owing to their contrasting stimuli-responsive fluorescence in solution along with strong emission in aggregated and solidstates, have been employed in optoelectronic devices, sensors, and bioimaging. Pyrene derivatives usually exhibit strong fluorescence and concentration-dependent excimer/aggregate emission in solution. However, the impacts of microenvironments on the monomer and aggregate emission bands and their relative intensities in solution, solid, and supramolecular aggregates are intriguing. The present study delineates a trade-off between the monomer and aggregate emissions of a pyrene-benzophenone derivative (ABzPy) in solution, in the solid-state, and in nanoaggregates through a combined spectroscopic and microscopic approach. The impact of external stimuli (viscosity, pH) on the aggregate emission was demonstrated using steady-state and time-resolved spectroscopy, including fluorescence correlation spectroscopy and fluorescence anisotropy decay analysis. The aggregate formation was noticed at a higher concentration (>10 µM) in solution, at 77 K (5 µM), and in the solid-state due to the π-π stacking interactions (3.6 Å) between two ABzPy molecules. In contrast, no aggregate formation was observed in the viscous medium as well as in a micellar environment even at a higher concentration of ABzPy (50 µM). The crystal structure analysis further shed light on the intermolecular hydrogen-bonding-assisted solid-state emission, which was found to be highly sensitive toward external stimuli like pH and mechanical forces. The broad emission band comprising both monomer and aggregate in the aqueous dispersion of nanoaggregates was used for the specific cellular imaging of lysosomes and lipid droplets, respectively.

Load transfer across a mandible during a mastication cycle: The effects of odontogenic tumour.

The extent to which load transfer in a diseased mandible with odontogenic tumour might influence the potential risk of pathological fracture has scarcely been investigated. The study sought to investigate the quantitative deviations in load transfer across healthy and cancer-affected (diseased) mandibles having odontogenic tumours. The effect of size of the tumours (small: 9 mm diameter, large: 19 mm diameter), and variation in bone mechanical (elastic) properties of the mandible on load transfer in cancer-affected mandibles during a mastication cycle have been investigated. Based on patient-specific computed tomography-scan datasets, detailed three-dimensional finite element models of healthy and diseased mandibles were developed. High stresses of 25-30 MPa and strains ∼700 µÎµ were observed in the healthy mandible during the right molar bite. However, marginal deviations were observed in principal stress distributions in the diseased mandibles with small- and large-sized tumours, as compared to the healthy mandible. Maximum principal strains of ∼1474 µÎµ were found in the body region adjacent to the symphysis region for small-sized tumour. Whereas for large-sized tumour, maximum strains of ∼2700 µÎµ were observed in the right buccal regions. Reduction in Young's modulus due to different stages of odontogenic tumours had a localised effect on the principal stress distributions, but triggered an abrupt increase in the principal tensile strains. It appears that there is a potential risk of pathological fracture for large-sized odontogenic tumour, owing to high tensile stresses and strains.

Design of porous titanium scaffold for complete mandibular reconstruction: The influence of pore architecture parameters.

Patients having a medical history of oral cancer, infectious diseases or trauma are often advised surgical intervention with customized complete mandibular constructs (CMC) made of Titanium (Ti) scaffolds. A numerical framework based on a homogenization technique was developed to investigate the influence of pore architecture parameters on homogenized orthotropic material properties of the scaffolds. A comparative 3D Finite Element (FE) analysis of six CMC models, having homogenized orthotropic material properties, under a mastication cycle, was undertaken to pre-clinically determine the optimal CMC for a patient. Orthotropic material properties of Ti-scaffolds decreased with an increase in the inter-strut distance. Stress and strain distributions of CMC models during right molar bite were investigated. Despite small differences in stress distributions in the 'body' region of CMC models, the overall stress distribution (tensile and compressive) of CMC models (30-32 MPa) were well comparable to that of an intact mandible (34.54 MPa). Higher magnitudes of tensile strains were observed for models with 0.2 mm (9884µÉ›) and 0.4 mm strut diameter (SD), both having 0.5 mm inter-strut distance (ID), at articular condyle area, body and symphysis equivalent part of the constructs. The maximum principal tensile strains were higher in the CMC models with 0.5 mm ID as compared to those having 0.3 mm ID. Comparatively, the scaffolds with lesser ID (0.3 mm) resulted in higher stiffness, thereby evoking less principal strains in the CMC models. Moreover, considering the weight of the scaffolds, the CMC models having 0.3 mm ID with 0.2 mm SD and 0.5 mm ID with 0.6 mm SD seemed most appropriate for a patient.

Doping of carbon nanodots for saving cells from silver nanotoxicity: A study on recovering osteogenic differentiation potential.

Silver nanoparticles are explored for many advanced biological applications including the development of antimicrobial surfaces on implants, SERS imaging, nanotherapeutics, biosensing and much more. However, recent research findings suggest silver nanoparticles provide blockade of differentiation of mesenchymal stem cells (MSCs), especially into osteogenic developmental pathway via generation of reactive oxygen species. These studies suggest that the application of silver nanoparticles in medical implants should be prohibited. In the current study, carbon nanodots (CND) supported silver clusters (AgC) is explored as a remedy to this problem. The nanostructure was synthesized in microwave irradiation induced rapid method and characterization was conducted via UV-Vis spectroscopy, fluorescence spectroscopy, HRTEM, XRD, FTIR, Raman spectroscopy, DLS, AFM, and XPS. Fluorescence spectrum showed a quantum yield of 0.25 while Raman spectroscopy showed rapid amplification of CND specific peaks implicating significant SERS property. Further in vitro biocompatibility (MTT) and bio-imaging capability was assessed culturing Wharton's Jelly-derived MSCs. In this study, its efficacy as in-situ cellular oxidative stress scavenger is also studied using NBT and DCFH-DA assay. Via ALP assay, alizarin red staining, cell membrane nanoindentation studies, PCR analysis and immunocytochemistry for osteoblast-like gene expression it was confirmed that AgCs can control silver nanoparticle-induced inhibition of osteogenic differentiation in vitro. Thus, AgCs (Carbon nanodots supported silver clusters) are not only considered to be a dual-mode bio-imaging nanoprobe but also a remedy to the silver-induced ROS generation and osteogenic differentiation blockade of MSCs.

Carbon Nanodots Doped Super-paramagnetic Iron Oxide Nanoparticles for Multimodal Bioimaging and Osteochondral Tissue Regeneration via External Magnetic Actuation.

Super-paramagnetic iron oxide nanoparticles (SPIONs) have multiple theranostics applications such as T2 contrast agent in magnetic resonance imaging (MRI) and electromagnetic manipulations in biomedical devices, sensors, and regenerative medicines. However, SPIONs suffer from the limitation of free radical generation, and this has a certain limitation in its applicability in tissue imaging and regeneration applications. In the current study, we developed a simple hydrothermal method to prepare carbon quantum dots (CD) doped SPIONs (FeCD) from easily available precursors. The nanoparticles are observed to be cytocompatible, hemocompatible, and capable of scavenging free radicals in vitro. They also have been observed to be useful for bimodal imaging (fluorescence and MRI). Further, 3D printed gelatin-FeCD nanocomposite nanoparticles were prepared and used for tissue engineering using static magnetic actuation. Wharton's jelly derived mesenchymal stem cells (MSCs) were cultured on them with magnetic actuation and implanted at the subcutaneous region. The tissues obtained have shown features of both osteogenic and chondrogenic differentiation of the stem cells in vivo. In vitro, PCR studies show MSCs express gene expression of both bone and cartilage-specific markers, suggesting FeCDs under magnetic actuation can lead MSCs to go through differentiating into an endochondral ossification route.

Core-Shell Nanofibrous Scaffold Based on Polycaprolactone-Silk Fibroin Emulsion Electrospinning for Tissue Engineering Applications.

The vast domain of regenerative medicine comprises complex interactions between specific cells' extracellular matrix (ECM) towards intracellular matrix formation, its secretion, and modulation of tissue as a whole. In this domain, engineering scaffold utilizing biomaterials along with cells towards formation of living tissues is of immense importance especially for bridging the existing gap of late; nanostructures are offering promising capability of mechano-biological response needed for tissue regeneration. Materials are selected for scaffold fabrication by considering both the mechanical integrity and bioactivity cues they offer. Herein, polycaprolactone (PCL) (biodegradable polyester) and 'nature's wonder' biopolymer silk fibroin (SF) are explored in judicious combinations of emulsion electrospinning rather than conventional electrospinning of polymer blends. The water in oil (W/O) emulsions' stability is found to be dependent upon the concentration of SF (aqueous phase) dispersed in the PCL solution (organic continuous phase). The spinnability of the emulsions is more dependent upon the viscosity of the solution, dominated by the molecular weight of PCL and its concentration than the conductivity. The nanofibers exhibited distinct core-shell structure with better cytocompatibility and cellular growth with the incorporation of the silk fibroin biopolymer.

Emblica officinalis Garten fruits extract ameliorates reproductive injury and oxidative testicular toxicity induced by chlorpyrifos in male rats.

Organophosphate pesticides have destroying properties on male reproduction and chlorpyrifos adversely affects the male reproductive system. Emblica offcinalis Garten plays a vital role to challenge many diseases in human body. We investigated the induction of oxidative stress in the male reproductive system of adult rats (Wistar Strain) exposed to widely used organophosphate pesticide, Chlorpyrifos, and tried to establish the ameliorative properties of Emblica officinalis Garten with respect to reproductive reconstruction in them. Rats were divided into 2 groups, control group and experimental group, and the experimental group was divided into 3 groups (G1-G3). All the groups had 5 rats each. Control group received water, experimental group, G1, received 20 mg/kg bw/day Emblica officinalis Garten, G2 received 12 mg/kg bw/day chlorpyrifos and G3 received 12 mg chlorpyrifos with 20 mg Emblica officinalis Garten /kg bw/day. Treatment was done orally from 30 days. Thereafter body weight, male reproductive organs weight, sperm count, sperm morphology, ACP, ALP, total protein, uric acid and testis and serum testosterone level were determined using standard methods. The changes recorded are indicative of infertility in male rats because of chlorpyrifos exposure. When the subjects were treated with Emblica officinalis Garten in conjunction with chlorpyrifos, these parameters exhibited recovery and when treated with Emblica officinalis Garten alone, these parameters were more or less near to the control group. This highlights the debilitating effect of chlorpyrifos and scavenging property of Emblica officinalis Garten.

Oryzias melastigma - an effective substitute for exotic larvicidal fishes: enhancement of its reproductive potential by supplementary feeding.

A preliminary study was conducted on the efficacy of Oryzias melastigma in consuming mosquito larva so as to control mosquito and mosquito borne diseases, and enhancing its reproductive success using supplementary feed. Oryzias melastigma is a larvivore fish and widely distributed in the shallow water, wetlands of Gangetic plains and peninsular India. These studies indicate that O. melastigma is a prolific breeder and gregarious feeder of mosquito larvae. Increased reproduction by providing different supplementary feed, of which Ulothrix acted remarkably, may aid in wide spread use of this fish as a biological control measure against mosquitoes. One adult fish of any sex can consume 87.1% first instars mosquito larvae/day. So, early stages of mosquito larvae are effectively controlled, as compared to other successive stages. Ulothrix has considerable effect on egg production, successful hatching and regaining reproductive maturity of female in surprisingly quicker interval.