Based on the self-assembly process occurring in the human body all the time, self-assembled nanomaterials were designed by the researchers. The self-assembled nanomaterials have controllability, biocompatibility and functional advantages in vivo. The self-assembled nanomaterials constructed in situ under a physiological environment display various biological characteristics which can be used for imaging, therapy, and broad clinical applications. In situ self-assembled nanomaterials can boost drug function, reduce toxic and side effects, prolong imaging time and enlarge signal-to-noise ratio. By using pathological conditions to trigger specific responses in vivo, well-ordered nanoaggregates can be spontaneously formed by multiple weak bonding interactions. The assembly shows higher accumulation and longer retention in situ. Endogenous triggers for in situ assembly, such as enzymes, pH, reactive oxygen species and ligand receptor interaction, can be used to transform the materials into a variety of controllable nanostructures including nanoparticles, nanofibers and gels through bioactivated in vivo assembly (BIVA) strategies. BIVA strategies can be applied for treatment, imaging or participate in the physiological activities of cells at the lesion site. This review summarized and prospected the design of self-assembled peptide materials based on BIVA technology and their biomedical applications. The nanostructures of the self-assembly enable some beneficial biological effects, such as assembly induced retention (AIR) effect, enhanced targeting effect, multivalent bond effect, and membrane disturbance. Thus, the BIVA nanotechnology is promising for efficient drug delivery, enhancement of targeting and treatment, as well as optimization of the biological distribution of drugs.
Drug Delivery Systems ; Humans ; Nanofibers/chemistry* ; Nanoparticles ; Nanostructures/chemistry* ; Peptides

Objective: To analysis the biomechanical and biocompatible properties of calcium phosphate cement (CPC) enhanced by chitosan short nanofibers(CSNF) and Arg-Gly-Asp (RGD). Methods: Chitosan nanofibers were prepared by electrospinning, and cut into short fibers by high speed dispersion. CPC with calcium phosphorus ratio of 1.5:1 was prepared by Biocement D method. The composition and structure of CPC, CSNF, RGD modified CSNF (CSNF-RGD), CSNF enhanced CPC (CPC-CSNF), RGD modified CPC-CSNF (CPC-CSNF-RGD) were observed by infrared spectrum, X-ray diffraction (XRD) and scan electron microscopy (SEM). The mechanical properties were measured by universal mechanical testing instrument. The adhesion and proliferation of MC3T3 cells were assessed using immunofluorescence staining and MTT method. Results: The distribution of CSNF in the scaffold was homogeneous, and the porous structure between the nanofibers was observed by SEM. The infrared spectrum showed the characteristic peaks at 1633 nm and 1585 nm, indicating that RGD was successfully grafted on chitosan nanofibers. The XRD pattern showed that the bone cement had a certain curability. The stain-stress test showed that break strengths were (17.74±0.54) MPa for CPC-CSNF and (16.67±0.56) MPa for CPCP-CSNF-RGD, both were higher than that of CPC(all P<0.05). The immunofluorescence staining and MTT results indicated that MC3T3 cells grew better on CPC-CSNF-RGD after 240 min of culture(all P<0.05). Conclusion: CSNF-RGD can improve the biomechanical property and biocompatibility of CPC, indicating its potential application in bone tissue repair.
3T3 Cells ; Animals ; Biocompatible Materials ; Bone Cements ; chemistry ; metabolism ; pharmacology ; Calcium Phosphates ; metabolism ; Cell Proliferation ; drug effects ; Chitosan ; chemistry ; pharmacology ; Mice ; Nanofibers ; chemistry ; Oligopeptides ; chemistry

Since the release rate of protein in hydrogels is directly dependent upon the size of the protein and the hydrogel, how to deliver low molecular weight protein for prolonged periods has always been a problem. In this article, we present a usage of self-assembling peptide (P3) with the RGD epitope on its N terminus. The concentration of the released insulin-like growth factor 1 (IGF-1) was determined by UV-vis spectroscopy and the release kinetics suggested a notable reduction of the IGF-1 release rate. Cell entrapment experiments revealed that IGF-1 delivery by biotinylated nanofibers could promote the proliferation of the mouse chondrogenic ATDC5 cells when compared with cells embedded within nanofibers with untethered IGF-1.
Animals ; Biotin ; Cell Line ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Hydrogels ; chemistry ; Insulin-Like Growth Factor I ; pharmacology ; Mice ; Nanofibers ; Oligopeptides ; chemistry

Amyloid fibrils belong to a category of abnormal aggregations of natural proteins, which are closely related to many human diseases. Recently, some critical peptide sequences have been extensively studied for clarifying the molecular mechanism of natural proteins to form amyloid fibrils. In the present study, we designed a short peptide GGAAVV (GAV-6) composed of hydrophobic amino acids glycine (G), alanine (A) and valine (V) and studied its ability to form amyloid fibrils. As characterized by atomic force microscopy (AFM) and dynamic light scattering (DLS), the peptide could self-assemble into smooth nanofibers without branches. Congo red staining/binding and thioflavin-T (ThT) binding experiments show that the nanofibers formed by GAV-6 shared identical properties with typical amyloid fibrils. These results show that the designed peptide GAV-6 could self-assemble into typical amyloid fibrils, which might make it a useful model molecule to clarify the mechanism for the formation of amyloid fibrils in the future.
Amino Acid Sequence ; Amyloid ; chemistry ; Humans ; Microscopy, Atomic Force ; Models, Molecular ; Nanofibers ; chemistry ; Peptides ; chemistry

OBJECTIVE: To investigate and compare the growth of human fetal retinal pigment epithelial (RPE) cells seeded onto electrospun polyamide nanofibers (EPN) or etched pore polyester (EPP), and, further, to explore their possible use as prosthetic Bruch's membrane. METHODS: Human fetal RPE cells were planted onto the EPN, EPP and plastic (control) substrates in Transwells. The cultures were assessed with respect to cell attachment at 2, 4, 8 hours and proliferation at 1, 4, 8 days after seeding. Growth and morphology of the cells were monitored under the phase contrast microscope, and the phenotype was identified by immunofluorescence staining with antibodies against tight junction protein ZO-1. Strips of single EPP coated with nothing or EPP coated with EPN was differently implanted into the subretinal space of two P21 RCS rats for two weeks and the histologic slides of the retina were assessed. RESULTS: Cultured human fetal RPE cells were attached to either EPN or EPP substrates (with seeding on plastic substrate as control). After 8 h, the numbers of adherent cells in the EPN, EPP and control groups were 1.23*10(5)/cm(2), 1.70*10(5)/cm(2), and 1.64*10(5)/cm(2), respectively. The number of RPE cells attached to EPN was obviously less than that to both EPP and control (P<0.05). On the first day, the proliferation of cells on EPN was less than that of EPP and control (P<0.05); but by the 8th day in culture, the proliferation of cells on EPN had increased and was higher than proliferation on both EPP and control (P<0.05). All of the RPE cells cultured on EPN and EPP substrates were in monolayer, and the EPN-attached cells resembled the inner collagenous layer of Bruch's membrane. Immunofluorescence staining showed that the RPE cells cultured on EPN and EPP substrates adopted a higher expression of ZO-1 than that on the plastic control substrate. Subretinal implantation of either EPP alone or EPP as a carrier for free EPN for 2 weeks in P21RCS rats resulted in an expected encapsulation and loss of photoreceptor layer. No toxicity or other adverse reaction was observed in the vicinity of the transplant. CONCLUSION EPN and EPP could maintain human fetal RPE cell attachment and proliferation. Both EPN and EPP appeared to be grossly tolerance and biocompatible with subretinal implantation. EPN represents an intriguing prospect for prosthetic Bruch's membrane replacement because of its similarity in structure to native Bruch's membrane.
Animals ; Biocompatible Materials ; chemistry ; Bruch Membrane ; Cell Proliferation ; Cells, Cultured ; Fetus ; Humans ; Membranes, Artificial ; Nanofibers ; chemistry ; Polyesters ; chemistry ; Porosity ; Rats ; Retinal Pigment Epithelium ; cytology ; growth & development ; Tissue Engineering

Peptide RADA can undergo spontaneous assembly into well ordered nanofibers and hydrogels in its water solution. In this work, a variety of proteins, including IGF-1, aFGF and VEGF with different molecular weight and isoelectric points, were chosen and encapsulated within the RADA peptide hydrogel. UV-vis spectroscopy was used to determine the concentration of the released proteins in the solution. The release kinetics suggested that protein diffusion through nanofiber hydrogels depended primarily on the size of the protein and the density of the peptide nanofiber. Circular dichroism (CD) spectroscopy indicated that the encapsulation and release by RADA hydrogel did not affect the secondary structure of the proteins studied.
Circular Dichroism ; Delayed-Action Preparations ; Fibroblast Growth Factor 1 ; metabolism ; Hydrogels ; chemistry ; Insulin-Like Growth Factor I ; metabolism ; Nanofibers ; Peptides ; chemistry ; Protein Structure, Secondary ; Vascular Endothelial Growth Factor A ; metabolism

Three-dimensional poly (epsilon-caprolactone)/silk sericin (PCL/SS) porous nanofibrous scaffolds were prepared by electrospinning. The structure and properties of the scaffolds were characterized by Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM), Fourier Transform Infrared Spectroscopy (FTIR) and water contact angle instrument. Studies on cell adhension and proliferation were carried out by culturing human primary skin fibroblast cells (FEK4) on these scaffolds using SEM and MTS. The experimental results showed that the PCL/SS nanofibrous scaffolds with SS nanoparticles had porous non-woven mesh structure with nanofibrous cross-linked with each other. Fiber diameter was very uniform and precise, and the secondary structure of SS protein had not been changed. Furthermore, the capability of hydrophile increased with the SS addition, which improved FEK4 cells adhesion and proliferation on the scaffolds.
Biocompatible Materials ; chemistry ; Cell Adhesion ; drug effects ; Cells, Cultured ; Fibroblasts ; cytology ; Microscopy, Electron ; Nanofibers ; chemistry ; Polyesters ; chemistry ; Sericins ; chemistry ; Silk ; chemistry ; Spectroscopy, Fourier Transform Infrared ; Tissue Scaffolds ; chemistry

Electrospun nanofibrous materials are considered as ideal scaffolds for tissue engineering because their fibrous structure is quite similar to the morphology of natural extracellular matrix, and they can offer biomimetic microenvironment for cell growth. However, the highly porous scaffolds are too weak to meet the mechanical requirement of guided tissue regeneration (GTR). In the present investigation, electrospun L-polylactic acid (PLLA) nanofibrous membranes were collected with high speed rolling method, and then hot stretched and annealed to improve the tensile strength and cell occlusivity. The membrane with the maximum tensile strength (strength 103MPa and modulus 1.83GPa) was obtained by hot-stretching for twice at 100 degrees C and further annealed for 10min at the same temperature. Cytotoxicity test showed that the heat treated membrane supported well the attachment and growth of human periodontal ligament cells, but inhibited the cell proliferation. The cell occlusivity of the membrane was also significantly improved as the porosity decreased after heat treatment. It could be used as the enhancement interlayer of barrier materials for GTR.
Biocompatible Materials ; Cells, Cultured ; Guided Tissue Regeneration ; Hot Temperature ; Humans ; Lactic Acid ; chemistry ; Membranes, Artificial ; Nanofibers ; Polyesters ; Polymers ; chemistry ; Primary Cell Culture ; Stress, Mechanical ; Tensile Strength ; Tissue Engineering ; Tissue Scaffolds ; chemistry

The present research was aimed to explore the biocompatibility of IKVAV self-assembling peptide nanofiber scaffold with olfactory ensheathing cells (OECs) of rats. The OECs were seeded onto the surface of coverslips covered with IKVAV self-assembling peptide nanofiber scaffold hydrogel (2D culture system), and implanted within IKVAV self-assembling peptide nanofiber scaffold hydrogel (3D culture system), respectively. The adhesion, viability of OECs were observed with inverted microscope. Then the characteristics for survival and adhesion of cells by image processing were observed, and statistical analysis on the number of S-100 positive cell, the area of the cell bodies and the perimeter of the cell and MTT method were carried out. It was found that the OECs could survive and migrate in IKVAV self-assembling peptide nanofiber scaffold. The result of the cell MTT exam, of the shape and quantity of cells had no significant difference compared to those of the OECs cultured with poly-L-lysine (PLL). It has been proved that IKVAV self-assembling peptide nanofiber scaffold has good biocompatibility with rat OECs.
Animals ; Animals, Newborn ; Biocompatible Materials ; chemistry ; Cell Proliferation ; Cells, Cultured ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Laminin ; chemistry ; Nanofibers ; chemistry ; Olfactory Bulb ; cytology ; drug effects ; Peptide Fragments ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

BACKGROUNDPeripheral nerve regeneration across large gaps is clinically challenging. Scaffold design plays a pivotal role in nerve tissue engineering. Recently, nanofibrous scaffolds have proven a suitable environment for cell attachment and proliferation due to similarities of their physical properties to natural extracellular matrix. Poly(propylene carbonate) (PPC) nanofibrous scaffolds have been investigated for vascular tissue engineering. However, no reports exist of PPC nanofibrous scaffolds for nerve tissue engineering. This study aimed to evaluate the potential role of aligned and random PPC nanofibrous scaffolds as substrates for peripheral nerve tissue and cells in nerve tissue engineering.

METHODSAligned and random PPC nanofibrous scaffolds were fabricated by electrospinning and their chemical characterization were carried out using scanning electron microscopy (SEM). Dorsal root ganglia (DRG) from Sprague-Dawley rats were cultured on the nanofibrous substrates for 7 days. Neurite outgrowth and Schwann-cell migration from DRG were observed and quantified using immunocytochemistry and SEM. Schwann cells derived from rat sciatic nerves were cultured in electrospun PPC scaffold-extract fluid for 24, 48, 72 hours and 7 days. The viability of Schwann cells was evaluated by 3-[4,5-dimethyl(thiazol-2-yl)-2,5-diphenyl] tetrazolium bromide (MTT) assay.

RESULTSThe diameter of aligned and random fibers ranged between 800 nm and 1200 nm, and the thickness of the films was approximately 10 - 20 µm. Quantification of aligned fiber films revealed approximately 90% alignment of all fibers along the longitudinal axis. However, with random fiber films, the alignment of fibers was random through all angle bins. Rat DRG explants were grown on PPC nanofiber films for up to 1 week. On the aligned fiber films, the majority of neurite outgrowth and Schwann cell migration from the DRG extended unidirectionally, parallel to the aligned fibers. However, on the random fiber films, neurite outgrowth and Schwann cell migration were randomly distributed. A comparison of cumulative neurite lengths from cultured DRGs indicated that neurites grew faster on aligned PPC films ((2537.6 ± 987.3) µm) than randomly-distributed fibers ((493.5 ± 50.6) µm). The average distance of Schwann cell migration on aligned PPC nanofibrous films ((2803.5 ± 943.6) µm) were significantly greater than those on random fibers ((625.3 ± 47.8) µm). The viability of Schwann cells cultured in aligned PPC scaffold extract fluid was not significantly different from that in the plain DMEM/F12 medium at all time points after seeding.

CONCLUSIONSThe aligned PPC nanofibrous film, but not the randomly-oriented fibers, significantly enhanced peripheral nerve regeneration in vitro, indicating the substantial role of topographical cues in stimulating endogenous nerve repair mechanisms. Aligned PPC nanofibrous scaffolds may be a promising biomaterial for nerve regeneration.

Animals ; Biocompatible Materials ; chemistry ; Cells, Cultured ; Ganglia, Spinal ; cytology ; metabolism ; ultrastructure ; Immunohistochemistry ; Microscopy, Electron, Scanning ; Nanofibers ; chemistry ; Nerve Regeneration ; physiology ; Nerve Tissue ; cytology ; metabolism ; ultrastructure ; Polymers ; chemistry ; Propane ; analogs & derivatives ; chemistry ; Rats ; Rats, Sprague-Dawley ; Schwann Cells ; cytology ; metabolism ; ultrastructure ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry