Sea squirt-inspired bio-derived tissue sealants.

Sea squirts' or tunicates' bodies are composed of cellulose nanofibers and gallol- functionalized proteins. These sea creatures are known to heal their injuries under seawater by forming crosslinks between gallols and functional groups from other proteins in their bodies. Inspired by their wound healing mechanism, herein, we have developed a tissue sealant using zein (a plant-based protein) and tannic acid (gallol-containing polyphenol). Except for fibrin- based sealants, most commercial surgical adhesives, and sealants available today are derived from petroleum products that compromise their biodegradability. They often have complicated and multi-step synthesis processes that ultimately affect their affordability. To overcome this challenge, we ensured that these sea squirt-inspired tissue sealants are bio-based, easily synthesized, and low-cost. The sealants were studied on their own and with a food-grade enzyme transglutaminase. The adhesion performances of the sealants were found to be higher than physiological pressures in seven out of nine different tissue substrates studied here. Their performance was also better than or on par with the FDA-approved fibrin sealant Tisseel. Ex vivo models demonstrate instant sealing of leaking wounds in less than a minute. The sealants were not only cytocompatible but also showed complete wound healing on par with sutures and Tisseel when applied in vivo on skin incisions in rats. Overall, these sea squirt-inspired bio-based sealants show great potential to replace currently available wound closure methods.

Effect of Cross-Linkers on Mussel- and Elastin-Inspired Adhesives on Physiological Substrates.

Surgical adhesives can be useful in wound closure because they reduce the risk of infection and pain associated with sutures and staples. However, there are no commercially available surgical adhesives for soft tissue wound closure. To be effective, soft tissue adhesives must be soft and flexible, strongly cohesive and adhesive, biocompatible, and effective in a moist environment. To address these criteria, we draw inspiration from the elasticity and resilience of elastin proteins and the adhesive of marine mussels. We used an elastin-like polypeptide (ELP) for the backbone of our adhesive material due to its elasticity and biocompatibility. A mussel-inspired adhesive molecule, l-3,4-dihydroxyphenylalanine (DOPA), was incorporated into the adhesive to confer wet-setting adhesion. In this study, an ELP named YKV was designed to include tyrosine residues and lysine residues, which contain amine groups. A modified version of YKV, named mYKV, was created through enzymatic conversion of tyrosine residues into DOPA. The ELPs were combined with iron(III) nitrate, sodium periodate, and/or tris(hydroxymethyl)phosphine (THP) cross-linkers to investigate the effect of DOPA- and amine-based cross-linking on adhesion strength and cure time on porcine skin in a warm, humid environment. Incorporation of DOPA into the ELP increased adhesive strength by 2.5 times and reduced failure rates. Iron cross-linkers improved adhesion in the presence of DOPA. THP increased adhesion for all proteins tested even in the absence of DOPA. Using multiple cross-linkers in a single formulation did not significantly improve adhesion. The adhesives with the highest performance (iron nitrate mixed with mYKV and THP mixed with YKV or mYKV) on porcine skin had 10-18 times higher adhesion than a commercial sealant and reached appreciable adhesive strength within 10 min.

pH-Sensitive Mechanical Properties of Elastin-Based Hydrogels.

Ionizable amino acids in protein-based hydrogels can confer pH-responsive behavior. Because elastin-like polypeptides (ELPs) have an established sequence and can crosslink to form hydrogels, they are an ideal system for creating pH-sensitive materials. This study examines different parameters that might affect pH-sensitive behavior and characterizes the mechanical and physical properties between pH 3 and 11 of three ELP-based crosslinked hydrogels. The first finding is that varying the amount of crosslinker affects the overall stiffness and resilience of the hydrogels but does not strongly affect water content, swelling ratio, or pH sensitivity. Second, the choice of two popular tag sequences, which vary in histidine and aspartic acid content, does not have a strong effect on pH-sensitive properties. Last, selectively blocking lysine and tyrosine residues through acetylation significantly decreases the pH-sensitive zeta potential. Acetylated hydrogels also demonstrate different behavior at low pH values with reduced swelling, reduced water content, and higher stiffness. Overall, this work demonstrates that ELP hydrogels with ionizable groups are promising materials for environmentally-responsive applications such as drug delivery, tissue engineering, and microfluidics.

Collagen Type I and II Blend Hydrogel with Autologous Mesenchymal Stem Cells as a Scaffold for Articular Cartilage Defect Repair.

Collagen type II is a promising material to repair cartilage defects since it is a major component of articular cartilage and plays a key role in chondrocyte function. This study investigated the chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (MSCs) embedded within a 3:1 collagen type I to II blend (Col I/II) hydrogel or an all collagen type I (Col I) hydrogel. Glycosaminoglycan (GAG) production in Col I/II hydrogels was statistically higher than that in Col I hydrogels or pellet culture, and these results suggested that adding collagen type II promoted GAG production. Col I/II hydrogels had statistically lower alkaline phosphatase (AP) activity than pellets cultured in a chondrogenic medium. The ability of MSCs encapsulated in Col I/II hydrogels to repair cartilage defects was investigated by creating two cartilage defects in the femurs of rabbits. After 13 weeks, histochemical staining suggested that Col I/II blend hydrogels provided favorable conditions for cartilage repair. Histological scoring revealed a statistically higher cartilage repair score for the Col I/II hydrogels compared to either the Col I hydrogels or empty defect controls. Results from this study suggest that there is clinical value in the cartilage repair capabilities of our Col I/II hydrogel with encapsulated MSCs.

Comparison between Catechol- and Thiol-Based Adhesion Using Elastin-like Polypeptides.

Different chemistries have been utilized for adhesive materials to achieve adhesion in a humidified environment. l-3,4-dihydroxyphenylalanine (DOPA) found in marine mussel adhesive proteins has generated great interest because DOPA participates in multiple reaction mechanisms that confer the ability to adhere in wet conditions. However, the mussel adhesive complex also contains proteins with a relatively high thiol content, and these proteins can contribute to adhesion through the formation of disulfide bonds or interactions with DOPA. This work probes the individual contributions and interactions of DOPA and thiol chemistries to adhesion. To do so, we took advantage of the sequence flexibility in elastin-like polypeptides (ELPs) to create model proteins with highly similar sequences that are rich in either DOPA or thiol residues. The sequence similarity between the two ELP adhesives allowed us to focus on the differences between DOPA- and thiol-based adhesion. Curing kinetics in a wet setting, capability to recover from disturbance in the curing process, and cytocompatibility of the two adhesives were compared. Both chemistries resulted in cytocompatible materials. However, thiol chemistry had faster curing kinetics and higher adhesion strengths, whereas DOPA chemistry showed better recovery from disturbances during the curing process. By utilizing both DOPA- and thiol-based chemistry simultaneously and adding iron ions, we achieved fast curing kinetics, strong adhesion strengths, and good recovery from disturbances to curing. These insights into the contribution of these chemistries to adhesion provide important lessons for researchers designing adhesives that work in a humid environment.

Incorporation of short, charged peptide tags affects the temperature responsiveness of positively-charged elastin-like polypeptides.

Elastin-like polypeptides (ELPs) are recombinant protein domains exhibiting lower critical solution temperature (LCST) behavior. This LCST behavior is controlled not only by intrinsic factors including amino acid composition and polypeptide chain length but also by non-ELP fusion domains. Here, we report that the presence of a composite non-ELP sequence that includes both His and T7 tags or a short Ser-Lys-Gly-Pro-Gly (SKGPG) sequence can dramatically change the LCST behavior of a positively-charged ELP domain. Both the His and T7 tags have been widely used in recombinant protein design to enable affinity chromatography and serve as epitopes for protein detection. The SKGPG sequence has been used to improve the expression of ELPs. Both the composite tag and the SKGPG sequence are <15% of the total length of the ELP fusion proteins. Despite the small size of the composite tag, its incorporation imparted pH-sensitive LCST behavior to the positively-charged ELP fusion protein. This pH sensitivity was not observed with the incorporation of the SKGPG sequence. The pH sensitivity results from both electrostatic and hydrophobic interactions between the composite tag and the positively-charged ELP domain. The hydrophobicity of the composite tag also alters the ELP interaction with Hofmeister salts by changing the overall hydrophobicity of the fusion protein. Our results suggest that incorporation of short tag sequences should be considered when designing temperature-responsive ELPs and provide insights into utilizing both electrostatic and hydrophobic interactions to design temperature-responsive recombinant proteins as well as synthetic polymers.

Redox-Responsive Resilin-Like Hydrogels for Tissue Engineering and Drug Delivery Applications.

Resilin, a protein found in insect cuticles, is renowned for its outstanding elastomeric properties. The authors' laboratory previously developed a recombinant protein, which consisted of consensus resilin-like repeats from Anopheles gambiae, and demonstrated its potential in cartilage and vascular engineering. To broaden the versatility of the resilin-like protein, this study utilizes a cleavable crosslinker, which contains a disulfide bond, to develop smart resilin-like hydrogels that are redox-responsive. The hydrogels exhibit a porous structure and a stable storage modulus (G') of ≈3 kPa. NIH/3T3 fibroblasts cultured on hydrogels for 24 h have a high viability (>95%). In addition, the redox-responsive hydrogels show significant degradation in a reducing environment (10 mm glutathione (GSH)). The release profiles of fluorescently labeled dextrans encapsulated within the hydrogels are assessed in vitro. For dextran that is estimated to be larger than the mesh size of the gel, faster release is observed in the presence of reducing agents due to degradation of the hydrogel networks. These studies thus demonstrate the potential of using these smart hydrogels in a variety of applications ranging from scaffolds for tissue engineering to drug delivery systems that target the intracellular reductive environments of tumors.

Critical factors for the bulk adhesion of engineered elastomeric proteins.

Many protein-based materials, such as soy and mussel adhesive proteins, have been the subject of scientific and commercial interest. Recently, a variety of protein adhesives have been isolated from diverse sources such as insects, frogs and squid ring teeth. Many of these adhesives have similar amino acid compositions to elastomeric proteins such as elastin. Although elastin is widely investigated for a structural biomaterial, little work has been done to assess its adhesive potential. In this study, recombinant elastin-like polypeptides were created to probe the factors affecting adhesion strength. Lap shear adhesion was used to examine the effects of both extrinsic factors (pH, concentration, cross-linker, humidity, cure time and cure temperature) and intrinsic factors (protein sequence, structure and molecular weight). Of the extrinsic factors tested, only humidity, cure time and cure temperature had a significant effect on adhesion strength. As water content was reduced, adhesion strength increased. Of the intrinsic factors tested, amino acid sequence did not significantly affect adhesion strength, but less protein structure and higher molecular weights increased adhesion strength directly. The strengths of proteins in this study (greater than 2 MPa) were comparable to or higher than those of two commercially available protein-based adhesives, hide glue and a fibrin sealant. These results may provide general rules for the design of adhesives from elastomeric proteins.

Integrin-Mediated Interactions Control Macrophage Polarization in 3D Hydrogels.

Adverse immune reactions prevent clinical translation of numerous implantable devices and materials. Although inflammation is an essential part of tissue regeneration, chronic inflammation ultimately leads to implant failure. In particular, macrophage polarity steers the microenvironment toward inflammation or wound healing via the induction of M1 and M2 macrophages, respectively. Here, this paper demonstrates that macrophage polarity within biomaterials can be controlled through integrin-mediated interactions between human monocytic THP-1 cells and collagen-derived matrix. Surface marker, gene expression, biochemical, and cytokine profiling consistently indicate that THP-1 cells within a biomaterial lacking cell attachment motifs yield proinflammatory M1 macrophages, whereas biomaterials with attachment sites in the presence of interleukin-4 (IL-4) induce an anti-inflammatory M2-like phenotype and propagate the effect of IL-4 in induction of M2-like macrophages. Importantly, integrin α2ß1 plays a pivotal role as its inhibition blocks the induction of M2 macrophages. The influence of the microenvironment of the biomaterial over macrophage polarity is further confirmed by its ability to modulate the effect of IL-4 and lipopolysaccharide, which are potent inducers of M2 or M1 phenotypes, respectively. Thus, this study represents a novel, versatile, and effective strategy to steer macrophage polarity through integrin-mediated 3D microenvironment for biomaterial-based programming.

A bioinspired elastin-based protein for a cytocompatible underwater adhesive.

The development of adhesives that can be applied and create strong bonds underwater is a significant challenge for materials engineering. When the adhesive is intended for biomedical applications, further criteria, such as biocompatibility, must be met. Current biomedical adhesive technologies do not meet these needs. In response, we designed a bioinspired protein system that shows promise to achieve biocompatible underwater adhesion coupled with environmentally responsive behavior that is "smart" - that is, it can be tuned to suit a specific application. The material, ELY16, is constructed from an elastin-like polypeptide (ELP) that can be produced in high yields from Escherichia coli and can coacervate in response to environmental factors such as temperature, pH, and salinity. To confer wet adhesion, we utilized design principles from marine organisms such as mussels and sandcastle worms. When expressed, ELY16 is rich in tyrosine. Upon modification with the tyrosinase enzyme to form mELY16, the tyrosine residues are converted to 3,4-dihydroxyphenylalanine (DOPA). Both ELY16 and mELY16 exhibit cytocompatibility and significant dry adhesion strength (>2 MPa). Modification with DOPA increases protein adsorption to glass and provides moderate adhesion strength (∼240 kPa) in a highly humid environment. Furthermore, this ELP exhibits a tunable phase transition behavior that can be formulated to coacervate in physiological conditions and provides a convenient mechanism for application underwater. Finally, mELY16 possesses significantly higher adhesion strength in dry, humid, and underwater environments compared with a commercially available fibrin sealant. To our knowledge, mELY16 provides the strongest bonds of any rationally designed protein when used completely underwater, and its high yields make it more viable for commercial application compared to natural adhesive proteins. In conclusion, this ELP shows great potential to be a new "smart" underwater adhesive.

Protein-engineered microenvironments can promote endothelial differentiation of human mesenchymal stem cells in the absence of exogenous growth factors.

Peripheral artery disease often requires treatments with vascular grafts for vessel reconstruction. Endothelialization of the vascular grafts is important to achieve long-term patency because endothelial cells regulate thrombosis, inflammation, and growth of smooth muscle cells. One potential source of endothelial cells is human mesenchymal stem cells (hMSCs), which can be routinely differentiated towards the endothelial lineage using exogenous growth factors such as vascular endothelial growth factor (VEGF). However, there are few studies that investigate the effect of materials on endothelial differentiation in the absence of growth factors. This study demonstrates that exogenous growth factors are not needed to achieve endothelial differentiation of hMSCs and that protein-based microenvironments promote endothelial differentiation. Specifically, we genetically engineered proteins containing a VEGF-mimicking peptide and resilin repeats and demonstrated that cells grown on the protein-engineered microenvironments were viable, had normal metabolic activity, and displayed increased endothelial-specific markers and endothelial function compared to negative control cells. In particular, cells cultured on our proteins formed networks that were statistically equivalent to positive control cells. We confirmed that the mere presence of protein on surfaces was insufficient to promote endothelial differentiation of hMSCs, and the specific composition of the RZ-QK protein appeared to be necessary for promoting differentiation. Thus, our protein-based materials are promising tools for obtaining endothelial cells for use in vascular grafts.

Enzymatic Cross-Linking of Resilin-Based Proteins for Vascular Tissue Engineering Applications.

Protein-based biomaterials have received significant attention for tissue engineering applications. For example, resilin-based protein gels have been produced with different cross-linking chemistries for applications in cartilage, cardiovascular, and vocal fold engineering. In this study, we investigate an alternative cross-linking chemistry to form resilin-based protein hydrogels and demonstrate the versatility of the gels for investigating cell response to matrix stiffness. Specifically, transglutaminase was used to cross-link proteins and resulted in gel surfaces more suitable for long-term cell attachment compared to those formed by a Mannich-type condensation reaction. Since matrix stiffness is an important determinant in modulating cell response, we first tuned matrix stiffness by varying total protein concentration. Next, we observed that matrix stiffness modulated cell spreading and endothelial differentiation of human mesenchymal stem cells. In particular, our results show that cells differentiated on our matrices, which have a stiffness similar to subendothelial layers, had statistically equivalent endothelial function compared to cells differentiated on hard glass surfaces. Thus, our protein-based matrix system is a promising tool to provide substrates favorable for long-term cell attachment and better mimics the native subendothelial environment compared to conventional hard culture substrates.

Modular protein domains: an engineering approach toward functional biomaterials.

Protein domains and peptide sequences are a powerful tool for conferring specific functions to engineered biomaterials. Protein sequences with a wide variety of functionalities, including structure, bioactivity, protein-protein interactions, and stimuli responsiveness, have been identified, and advances in molecular biology continue to pinpoint new sequences. Protein domains can be combined to make recombinant proteins with multiple functionalities. The high fidelity of the protein translation machinery results in exquisite control over the sequence of recombinant proteins and the resulting properties of protein-based materials. In this review, we discuss protein domains and peptide sequences in the context of functional protein-based materials, composite materials, and their biological applications.

Delivery strategies to control inflammatory response: Modulating M1-M2 polarization in tissue engineering applications.

Macrophages are key players in many physiological scenarios including tissue homeostasis. In response to injury, typically the balance between macrophage sub-populations shifts from an M1 phenotype (pro-inflammatory) to an M2 phenotype (anti-inflammatory). In tissue engineering scenarios, after implantation of any device, it is desirable to exercise control on this M1-M2 progression and to ensure a timely and smooth transition from the inflammatory to the healing stage. In this review, we briefly introduce the current state of knowledge regarding macrophage function and nomenclature. Next, we discuss the use of controlled release strategies to tune the balance between the M1 and M2 phenotypes in the context of tissue engineering applications. We discuss recent literature related to the release of anti-inflammatory molecules (including nucleic acids) and the sequential release of cytokines to promote a timely M1-M2 shift. In addition, we describe the use of macrophages as controlled release agents upon stimulation by physical and/or mechanical cues provided by scaffolds. Moreover, we discuss current and future applications of "smart" implantable scaffolds capable of controlling the cascade of biochemical events related to healing and vascularization. Finally, we provide our opinion on the current challenges and the future research directions to improve our understanding of the M1-M2 macrophage balance and properly exploit it in tissue engineering and regenerative medicine applications.

Surface density of vascular endothelial growth factor modulates endothelial proliferation and differentiation.

Therapeutic strategies aim to regulate vasculature either by encouraging vessel growth for tissue engineering or inhibiting vascularization around a tumor. Vascular endothelial growth factor (VEGF) is essential to these processes, and there are several strategies that manipulate VEGF signaling. Here we develop a method to control the surface density of VEGF, which is covalently attached to tissue culture polystyrene (TCPS), and explore cellular responses to surfaces with varying VEGF densities. We show that the crosslinker reduces but does not eliminate the biological activity of soluble VEGF as measured by endothelial proliferation. However, endothelial cells cultured on surfaces of covalently bound VEGF did not proliferate in response to surface cues. Interestingly, compared to cells incubated with soluble VEGF (10 ng/ml) and cultured on TCPS, lower cell proliferation was observed when endothelial cells were cultured on high VEGF surface densities (5.9 ng/cm(2)), whereas higher cell proliferation occurred when cells were cultured on low surface densities (0.04 ng/cm(2)). High density surfaces (5.9 ng/cm(2)) also acted in synergy with an inhibitor of VEGF receptors to further suppress endothelial cell proliferation. We also examined the effect of VEGF surfaces on endothelial differentiation of mesenchymal stem cells. No effect was observed when cells were cultured on VEGF surfaces; however, the VEGF surfaces acted in synergy with an inhibitor of VEGF receptors to decrease the ability of differentiated cells to form vascular networks. Together, these results suggest that surface density of bound VEGF can be used to modulate cell behavior and inhibit an angiogenic response.

Vascular endothelial growth factor does not accelerate endothelial differentiation of human mesenchymal stem cells.

For clinical applications of engineered vascular replacements, endothelial cells may not be available in sufficient quantities due to limited harvesting sites and slow in vitro expansion rates. Soluble vascular endothelial growth factor (VEGF) is often added to differentiate mesenchymal stem cells (MSCs) into endothelial cells; however, recent studies demonstrate that VEGF is not required to upregulate endothelial markers. In contrast to previous assumptions, this study demonstrates that exogenous VEGF does not enhance or accelerate the upregulation of common endothelial markers during endothelial differentiation of human MSCs. MSCs were cultured at confluence for up to 3 weeks in either basal medium or medium containing VEGF. Cells were examined for gene and protein expression as well as the ability to internalize acetylated low density lipoprotein. With either treatment, endothelial differentiation occurred as evidenced by upregulation of gene and protein expression of typical endothelial markers and the ability to internalize acetylated low density lipoproteins. Interestingly, the addition of VEGF at typical or high concentrations (50 or 100 ng/ml) did not result in differences in gene or protein expression levels of many typical endothelial markers. However, high concentrations of VEGF did significantly increase protein expression of the arterial marker Ephrin-B1. Thus, VEGF did not accelerate or enhance differentiation of human MSCs towards endothelial cells but was vital for specification of arterial fate.

Synthesis and characterization of recombinant abductin-based proteins.

Recombinant proteins are promising tools for tissue engineering and drug delivery applications. Protein-based biomaterials have several advantages over natural and synthetic polymers, including precise control over amino acid composition and molecular weight, modular swapping of functional domains, and tunable mechanical and physical properties. In this work, we describe recombinant proteins based on abductin, an elastomeric protein that is found in the inner hinge of bivalves and functions as a coil spring to keep shells open. We illustrate, for the first time, the design, cloning, expression, and purification of a recombinant protein based on consensus abductin sequences derived from Argopecten irradians . The molecular weight of the protein was confirmed by mass spectrometry, and the protein was 94% pure. Circular dichroism studies showed that the dominant structures of abductin-based proteins were polyproline II helix structures in aqueous solution and type II ß-turns in trifluoroethanol. Dynamic light scattering studies illustrated that the abductin-based proteins exhibit reversible upper critical solution temperature behavior and irreversible aggregation behavior at high temperatures. A LIVE/DEAD assay revealed that human umbilical vein endothelial cells had a viability of 98 ± 4% after being cultured for two days on the abductin-based protein. Initial cell spreading on the abductin-based protein was similar to that on bovine serum albumin. These studies thus demonstrate the potential of abductin-based proteins in tissue engineering and drug delivery applications due to the cytocompatibility and its response to temperature.

Investigating the effect of peptide agonists on the chondrogenic differentiation of human mesenchymal stem cells using design of experiments.

Human mesenchymal stem cells (MSCs) are attractive for use in cartilage tissue engineering. Cells are often seeded in a structural scaffold containing growth factors. Peptide mimics of full-length growth factors are a promising alternative because they are less expensive and easier to manufacture. We investigated four short peptides for their effect on the chondrogenesis of human MSCs. The peptides were originally designed to mimic bone morphogenetic protein-2 (BMP-2), transforming growth factor-beta 1 (TGF-ß1), and insulin, all of which have been shown to affect MSC chondrogenesis. Previous studies demonstrated that the peptides elicited bioactivity in other cell types, but the peptides have not been investigated for their effect on chondrogenesis in human MSCs. In a preliminary investigation, peptides were added to a pellet culture of human MSCs and assayed for their effect on glycosaminoglycan (GAG) production. These experiments determined peptide concentrations used in a full-factorial experiment to investigate any interactions. The experiment revealed the BMP peptide as a robust stimulant for GAG production. .

Characterization of resilin-based materials for tissue engineering applications.

Modular proteins have emerged as powerful tools in tissue engineering because both the mechanical and biochemical properties can be precisely controlled through amino acid sequence. Resilin is an attractive candidate for use in modular proteins because it is well-known for having low stiffness, high fatigue lifetime, and high resilience. However, no studies have been conducted to assess resilin's compressive properties, cytocompatibility with clinically relevant cells, or effect on cell spreading. We designed a modular protein containing repeating sequences of a motif derived from Anopheles gambiae and cell-binding domains derived from fibronectin. Rapid cross-linking with tris(hydroxymethyl)phosphine was observed. The hydrogels had a complex modulus of 22 ± 1 kPa and yield strain of 63%. The elastic modulus in compression, or unconfined compressive modulus, was 2.4 ± 0.2 MPa, which is on the same order as human cartilage. A LIVE/DEAD assay demonstrated that human mesenchymal stem cells cultured on the resilin-based protein had a viability of 95% after three days. A cell-spreading assay revealed that the cells interacted with the fibronectin-derived domain in a sequence-specific manner and resulted in a mean cell area ~1.4-fold larger than when cells were seeded on a sequence-scrambled negative control protein. These results demonstrate that our resilin-based biomaterial is a promising biomaterial for cartilage tissue engineering.

Bone morphogenetic protein-derived peptide promotes chondrogenic differentiation of human mesenchymal stem cells.

Directing chondrogenic differentiation of human mesenchymal stem cells (MSCs) is currently a challenging problem in tissue engineering of cartilage. Short-peptide motifs are promising new tools to aid in controlling chondrogenesis. The aim of this study was to investigate whether a short bone morphogenetic protein-2 (BMP-2)-derived peptide (BMP peptide) stimulates chondrogenesis of human MSCs in the absence of other growth factors. A high-throughput pellet culture system was used to rapidly collect biochemical data such as glycosaminoglycan (GAG), total collagen, and DNA content, as well as alkaline phosphatase (AP) activity. Cells cultured with ≥100 µg/mL of the peptide produced 74% of the GAG content that cells cultured with BMP-2 produced. Comparable levels of GAG production were promoted by the peptide and BMP-2 over 4 weeks of culture. However, histology revealed that the peptide promoted a more homogenous distribution of GAG than BMP-2. The BMP peptide directed human MSCs to increase collagen production after 3 weeks, but at significantly lower levels compared to BMP-2. Treatment with BMP-2 resulted in a large increase in hypertrophic markers such as AP activity and gene expression of type X collagen, whereas treatment with the peptide resulted in little-to-no increase in these markers. These results suggest that the BMP peptide could be an effective new tool for cartilage tissue engineering.