Preparation and evaluation of porous poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) hydroxyapatite composite scaffolds

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) and PHBHHx-hydroxyapatite (HAP) composite scaffolds have been prepared by phase separation and subsequent sublimation of the solvent for bone tissue engineering. Scanning electron microscopy (SEM), porosity measurement, mechanical tests, and thermogravimertric analysis (TGA) are used to analyze the physical properties of the scaffolds. The biocompatibility and osteoconductivity are assessed by examining the morphology, proliferation, and differentiation of MC3T3-E1 osteoprogenitor cells seeded on the scaffolds. The PHBHHx-HAP composite scaffolds show better mechanical properties, biocompatibility, and osteoconductivity than the PHBHHx scaffolds. The results suggest that PHBHHx-HAP composite scaffolds can be employed as a promising candidate for bone reconstruction.     

Fabrication and characterization of porous hyaluronic acid-collagen composite scaffolds

Hyaluronic acid (HA) plays a vital role in many tissues, influencing water content and mechanical function, and has been shown to have positive biological effects on cell behavior in vitro. To begin to determine whether these benefits can be accessed if HA is incorporated into collagen-based scaffolds for tissue engineering, HA-collagen composite matrices were prepared and selected properties evaluated. HA-collagen scaffolds were cross-linked with carbodiimide and loss rates of HA in culture medium assessed. Scaffold pore structures were evaluated by light and electron microscopy. Adult canine chondrocytes were grown in selected HA-collagen scaffolds to assess the effects of HA on cell behavior. Homogenous HA-collagen slurries were achieved when polyionic complexes were suppressed. HA was uniformly distributed through the scaffolds, which demonstrated honeycomb-like pores with interconnectivity among pores increasing as HA content increased. Virtually all of the HA added to the collagen slurry was incorporated into the composite scaffolds that underwent a 7-day cross-linking protocol. After 5 days in culture medium, the HA content in the scaffolds was 5-7% regardless of initial HA loading. After only 2 weeks in culture cartilaginous tissue was found in the chondrocyte-seeded HA-collagen scaffolds. This study contributes to the understanding of the effects of HA content, pH, and cross-link treatment on pore characteristics and degradation behavior essential for the design of HA-collagen scaffolds. The demonstration that these scaffolds can be populated by chondrocytes and support in vitro formation of cartilaginous tissue warrants further investigation of this material system for tissue engineering.     

Biocompatibility and osteogenesis of biomimetic Bioglass-Collagen-Phosphatidylserine composite scaffolds for bone tissue engineering

A novel biomimetic composite scaffold Bioglass-Collagen-Phosphatidylserine (BG-COL-PS) was fabricated with a freeze-drying technique. The macrostructure and morphology as well as mechanical strength of the scaffolds were characterized. Scanning electronic microscopy (SEM) showed that the BG-COL-PS scaffolds exhibited interconnected porous structures with pore sizes of several microns up to about 300 μm. The scaffolds have a porosity of 75.40% and the corresponding compressive strength of 1.5469 Mpa. Rat mesenchymal stem cells (rMSCs) were seeded on BG-COL-PS or BG-COL scaffolds and cultured for 21 days in vitro. Based on the results of SEM, dsDNA content, alkaline phosphatase (ALP) activity, osteogenic gene expression analysis and alizarin red staining, the responses of MSCs to the scaffold exhibited a higher degree of attachment, growth as well as osteogenic differentiation than those on BG-COL scaffolds in vitro. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both pure BG-COL-PS scaffolds and MSC/scaffold constructs were implanted in rat femurs defects for 6 weeks and studied histologically and radiographically. The in vivo results showed that BG-COL-PS composite scaffolds exhibited good biocompatibility and extensive osteoconductivity with host bone. Moreover, the BG-COL-PS/MSC constructs dramatically enhanced the efficiency of new bone formation than pure BG-COL-PS scaffolds or BG-COL/MSC constructs. All these results demonstrate the usefulness of PS composited BG-COL-PS scaffolds for inducing enhanced bone formation. The BG-COL-PS scaffolds fulfill the basic requirements of bone tissue engineering scaffold and have the potential to be applied in orthopedic and reconstructive surgery.     

Polyurethane biomaterials for fabricating 3D porous scaffolds and supporting vascular cells

Successful tissue engineering of vascular grafts largely depends on synthetic scaffolds that support the survival, proliferation, and differentiation of seeded cells. To investigate the utility of polyurethanes for vascular tissue engineering, three-dimensional porous polyurethane scaffolds with highly interconnected pore structures were fabricated by a pressure differential/particulate leaching technique. Ammonium chloride and paraffin porogens were prepared to fabricate the scaffolds. Grinding of ammonium chloride resulted in particulates with uniform particle sizes but irregular shapes. Paraffin particulates made by a dispersion method, on the other hand, had spherical shapes and uniform particle sizes. Polyurethane scaffolds fabricated from these particulates had open faced, highly interconnected channels that could allow cellular infiltration and nutrient delivery. Human coronary artery smooth muscle and endothelial cell interactions with polyurethane surfaces revealed these biomaterials to maintain the contractile phenotype of human coronary artery smooth muscle cells and the formation of endothelial monolayers. During longer culture times, surface modification with cell adhesive extracellular matrix (ECM) protein promoted vascular cell proliferation, maintenance of the differentiated phenotype and endothelial monolayer integrity. Our results suggest that these polyurethanes, in conjunction with cell adhesive ECM proteins, could also support vascular cells in three-dimensional bioreactor-based culture conditions.     

X-ray imaging optimization of 3D tissue engineering scaffolds via combinatorial fabrication methods

We have developed a combinatorial method for determining optimum tissue scaffold composition for several X-ray imaging techniques. X-ray radiography and X-ray microcomputed tomography enable non-invasive imaging of implants in vivo and in vitro. However, highly porous polymeric scaffolds do not always possess sufficient X-ray contrast and are therefore difficult to image with X-ray-based techniques. Incorporation of high radiocontrast atoms, such as iodine, into the polymer structure improves X-ray radiopacity but also affects physicochemical properties and material performance. Thus, we have developed a combinatorial library approach to efficiently determine the minimum amount of contrast agent necessary for X-ray-based imaging. The combinatorial approach is demonstrated in a polymer blend scaffold system where X-ray imaging of poly(desaminotyrosyl-tyrosine ethyl ester carbonate) (pDTEc) scaffolds is improved through a controlled composition variation with an iodinated-pDTEc analog (pI(2)DTEc). The results show that pDTEc scaffolds must include at least 9%, 16%, 38% or 46% pI(2)DTEc (by mass) to enable effective imaging by microradiography, dental radiography, dental radiography through 0.75cm of muscle tissue or microcomputed tomography, respectively. Only two scaffold libraries were required to determine these minimum pI(2)DTEc percentages required for X-ray imaging, which demonstrates the efficiency of this new combinatorial approach for optimizing scaffold formulations.     

Preparation and histological evaluation of biomimetic three-dimensional hydroxyapatite/chitosan-gelatin network composite scaffolds

A novel biodegradable hydroxyapatite/chitosan-gelatin network (HA/CS-Gel) composite of similar composition to that of normal human bone was prepared as a three-dimensional biomimetic scaffold by phase separation method for bone tissue engineering. Changing the solid content and the compositional variables of the original mixtures allowed control of the porosities and densities of the scaffolds. The HA granules were dispersed uniformly in the organic network with intimate interface contact via pulverizing and ultrasonically treating commercial available HA particles. Scaffolds of 90.6% porosity were used to examine the proliferation and functions of the cells in this three-dimensional microenvironment by culturing neonatal rat caldaria osteoblasts. Histological and immunohistochemical staining and scanning electron microscopy observation indicated that the osteoblasts attached to and proliferated on the scaffolds. Extracellular matrices including collagen I and proteoglycan-like substrate were synthesized, while osteoid and bone-like tissue formed during the culture period. Furthermore, the cell/scaffold constructs had good biomineralization effect after 3 weeks in culture.     

Microfabrication of complex porous tissue engineering scaffolds using 3D projection stereolithography

The success of tissue engineering will rely on the ability to generate complex, cell seeded three-dimensional (3D) structures. Therefore, methods that can be used to precisely engineer the architecture and topography of scaffolding materials will represent a critical aspect of functional tissue engineering. Previous approaches for 3D scaffold fabrication based on top-down and process driven methods are often not adequate to produce complex structures due to the lack of control on scaffold architecture, porosity, and cellular interactions. The proposed projection stereolithography (PSL) platform can be used to design intricate 3D tissue scaffolds that can be engineered to mimic the microarchitecture of tissues, based on computer aided design (CAD). The PSL system was developed, programmed and optimized to fabricate 3D scaffolds using gelatin methacrylate (GelMA). Variation of the structure and prepolymer concentration enabled tailoring the mechanical properties of the scaffolds. A dynamic cell seeding method was utilized to improve the coverage of the scaffold throughout its thickness. The results demonstrated that the interconnectivity of pores allowed for uniform human umbilical vein endothelial cells (HUVECs) distribution and proliferation in the scaffolds, leading to high cell density and confluency at the end of the culture period. Moreover, immunohistochemistry results showed that cells seeded on the scaffold maintained their endothelial phenotype, demonstrating the biological functionality of the microfabricated GelMA scaffolds.     

基于三维打印技术的多药控释型载药人工骨生物学评价

目的 评价三维打印技术制备的多药控释型载药人工骨的生物相容性.方法 利用三维打印技术制备多药控释型载药人工骨,并进行急性毒性试验、热源试验、皮肤刺激试验、溶血试验、微核试验和肌肉埋植试验等检测评价其生物相容性.结果 该多药控释型载药人工骨无全身急性毒性反应、无热源效应;皮肤刺激实验局部皮肤未见红斑、水肿反应;溶血率为0.29%,有良好的血液相容性;微核实验未见致突变现象.无细胞遗传毒性作用;肌肉埋植实验未见局部组织变性、坏死或排斥现象.结论 三维打印技术制备的多药控释型载药人工骨具有良好的生物相容性,符合医用生物材料的性能要求. Abstract： Objective To evaluate the biocompatibility of porous drug implant scaffolds prepared by 3D printing technique. Methods Porous drug implant scaffolds were fabricated by 3D printing technique, and a series of tests were carried out to validate the biocompatibility, including acute systemic toxicity, hot source test, local irritation reaction, micronucleus test, muscle implant test and so on. Results The porous drug implant scaffolds showed no acute systemic toxicity, no pyrogenetic effect, no local erythema and edema in local irritation re-action, hemolysis rate of 0.29%, no cellular genetoxic. No local tissue denaturation, necrosis and exclusion were found in intramuscular implant test. Conclusion With good biocompatibility, the porous drug implant scaffold fabricated by 3D printing technique can meet the clinical requirement for biomaterial.     

Design and fabrication of porous biodegradable scaffolds: a strategy for tissue engineering

Current strategies of tissue engineering are focused on the reconstruction and regeneration of damaged or deformed tissues by grafting of cells with scaffolds and biomolecules. Recently, much interest is given to scaffolds which are based on mimic the extracellular matrix that have induced the formation of new tissues. To return functionality of the organ, the presence of a scaffold is essential as a matrix for cell colonization, migration, growth, differentiation and extracellular matrix deposition, until the tissues are totally restored or regenerated. A wide variety of approaches has been developed either in scaffold materials and production procedures or cell sources and cultivation techniques to regenerate the tissues/organs in tissue engineering applications. This study has been conducted to present an overview of the different scaffold fabrication techniques such as solvent casting and particulate leaching, electrospinning, emulsion freeze-drying, thermally induced phase separation, melt molding and rapid prototyping with their properties, limitations, theoretical principles and their prospective in tailoring appropriate micro-nanostructures for tissue regeneration applications. This review also includes discussion on recent works done in the field of tissue engineering.     

Biocompatibility and osteogenesis of biomimetic nano-hydroxyapatite/polyamide composite scaffolds for bone tissue engineering

In this study, we prepared nano-hydroxyapatite/polyamide (n-HA/PA) composite scaffolds utilizing thermally induced phase inversion processing technique. The macrostructure and morphology as well as mechanical strength of the scaffolds were characterized. Mesenchymal stem cells (MSCs) derived from bone marrow of neonatal rabbits were cultured, expanded and seeded on n-HA/PA scaffolds. The MSC/scaffold constructs were cultured for up to 7 days and the adhesion, proliferation and differentiation of MSCs into osteoblastic phenotype were determined using MTT assay, alkaline phosphatase (ALP) activity and collagen type I (COL I) immunohistochemical staining and scanning electronic microscopy (SEM). The results confirm that n-HA/PA scaffolds are biocompatible and have no negative effects on the MSCs in vitro. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both pure n-HA/PA scaffolds and MSC/scaffold constructs were implanted in rabbit mandibles and studied histologically and microradiographically. The results show that n-HA/PA composite scaffolds exhibit good biocompatibility and extensive osteoconductivity with host bone. Moreover, the introduction of MSCs to the scaffolds dramatically enhanced the efficiency of new bone formation, especially at the initial stage after implantation. In long term (more than 12 weeks implantation), however, the pure scaffolds show as good biocompatibility and osteogenesis as the hybrid ones. All these results indicate that the scaffolds fulfill the basic requirements of bone tissue engineering scaffold, and have the potential to be applied in orthopedic, reconstructive and maxillofacial surgery.     

3D printing of composite calcium phosphate and collagen scaffolds for bone regeneration

Low temperature 3D printing of calcium phosphate scaffolds holds great promise for fabricating synthetic bone graft substitutes with enhanced performance over traditional techniques. Many design parameters, such as the binder solution properties, have yet to be optimized to ensure maximal biocompatibility and osteoconductivity with sufficient mechanical properties. This study tailored the phosphoric acid-based binder solution concentration to 8.75 wt% to maximize cytocompatibility and mechanical strength, with a supplementation of Tween 80 to improve printing. To further enhance the formulation, collagen was dissolved into the binder solution to fabricate collagen-calcium phosphate composites. Reducing the viscosity and surface tension through a physiologic heat treatment and Tween 80, respectively, enabled reliable thermal inkjet printing of the collagen solutions. Supplementing the binder solution with 1–2 wt% collagen significantly improved maximum flexural strength and cell viability. To assess the bone healing performance, we implanted 3D printed scaffolds into a critically sized murine femoral defect for 9 weeks. The implants were confirmed to be osteoconductive, with new bone growth incorporating the degrading scaffold materials. In conclusion, this study demonstrates optimization of material parameters for 3D printed calcium phosphate scaffolds and enhancement of material properties by volumetric collagen incorporation via inkjet printing.     

Interactions of coronary artery smooth muscle cells with 3D porous polyurethane scaffolds

One strategy in vascular tissue engineering is the design of hybrid vascular substitutes where vascular cells infiltrate biostable porous scaffolds that provides favorable environment for guided cell repopulation and acts as a mechanically supporting layer after the tissue regeneration process. The aim of the present work was to study the interaction of human coronary artery smooth muscle cells (HCASMC) with 3D porous polyurethane scaffolds. We therefore fabricated porous and highly interconnected 3D polyurethane scaffolds that can promote HCASMC attachment, proliferation, and migration. SEM and microCT studies of the fabricated scaffolds showed that the current scaffolds had highly open and interconnected pore structures, with an average porosity of 84%. HCASMC interaction on polyurethane films revealed that cells adhere and express specific marker proteins (vinculin and h-caldesmon). This expression was further enhanced by coating the polyurethane with Matrigel. On uncoated 3D scaffolds, dense spherical aggregates of cells were often encountered with little adhesion of individual cells alongside the struts of the scaffold, independent of the porogens used. In contrast, when cultured on Matrigel-coated scaffolds, cell numbers quickly increased after 14 days and spread along the entire scaffold. At the upper scaffold surface, elongated cells were seen adhering to one another and also to the scaffold surface. These cells were elongated, aligned in parallel and contained abundant F-actin bundles suggesting a differentiated contractile phenotype. Deep into the scaffold, cells were encountered that formed actin-rich lamellipodial extensions spreading along the strut and lacked stress fibers, suggesting active cell migration along the substrate.     

In vitro analog of human bone marrow from 3D scaffolds with biomimetic inverted colloidal crystal geometry

In vitro replicas of bone marrow can potentially provide a continuous source of blood cells for transplantation and serve as a laboratory model to examine human immune system dysfunctions and drug toxicology. Here we report the development of an in vitro artificial bone marrow based on a 3D scaffold with inverted colloidal crystal (ICC) geometry mimicking the structural topology of actual bone marrow matrix. To facilitate adhesion of cells, scaffolds were coated with a layer of transparent nanocomposite. After seeding with hematopoietic stem cells (HSCs), ICC scaffolds were capable of supporting expansion of CD34+ HSCs with B-lymphocyte differentiation. Three-dimensional organization was shown to be critical for production of B cells and antigen-specific antibodies. Functionality of bone marrow constructs was confirmed by implantation of matrices containing human CD34+ cells onto the backs of severe combined immunodeficiency (SCID) mice with subsequent generation of human immune cells.     

Fabrication, characterization, and biocompatibility of single-walled carbon nanotube-reinforced alginate composite scaffolds manufactured using freeform fabrication technique

Composite polymeric scaffolds from alginate and single-walled carbon nanotube (SWCNT) were produced using a freeform fabrication technique. The scaffolds were characterized for their structural, mechanical, and biological properties by scanning electron microscopy, Raman spectroscopy, tensile testing, and cell-scaffold interaction study. Three-dimensional hybrid alginate/SWCNT tissue scaffolds were fabricated in a multinozzle biopolymer deposition system, which makes possible to disperse and align SWCNTs in the alginate matrix. The structure of the resultant scaffolds was significantly altered due to SWCNT reinforcement, which was confirmed by Raman spectroscopy. Microtensile testing presented a reinforcement effect of SWCNT to the mechanical strength of the alginate struts. Ogden constitutive modeling was utilized to predict the stress-strain relationship of the alginate scaffold, which compared well with the experimental data. Cellular study by rat heart endothelial cell showed that the SWCNT incorporated in the alginate structure improved cell adhesion and proliferation. Our study suggests that hybrid alginate/SWCNT scaffolds are a promising biomaterial for tissue engineering applications.     

Fabrication and characterization of porous alginate/polyvinyl alcohol hybrid scaffolds for 3D cell culture

Porous alginate/polyvinyl alcohol (PVA) hybrid scaffolds as bioartificial cell scaffolds were fabricated to improve cell compatibility as well as flexibility of the scaffolds. The alginate/PVA hybrid scaffolds with different PVA compositions up to 50 wt% were fabricated by a modified freeze-drying method including the physical cross-linking of PVA and the following chemical cross-linking of alginate. The prepared alginate/PVA hybrid scaffolds were characterized by morphology observations using scanning electron microscopy (SEM), the measurements of porosity and average pore sizes and the measurements of compressive strength and modulus. The scaffolds exhibited highly porous, open-cellular pore structures with almost the same surface and cross-sectional porosities (total porosities about 85%, regardless of PVA composition) and the pore sizes from about 290 microm to about 190 microm with increasing PVA composition. The alginate/PVA hybrid scaffolds were more soft and elastic than the control alginate scaffold without significant changes of mechanical strength. The scaffolds were examined for their in vitro cell compatibility by the culture of chondrocytes (human chondrocyte cell line) in the scaffolds and the following analyses by MTT assay and SEM observation. It was observed that the alginate/PVA scaffolds had better cell adhesion and faster growth than the control alginate scaffold. It seems that 30 wt% addition of PVA to alginate in the fabrication of the hybrid scaffolds is desirable for improving their flexibility and cell compatibility.     

3D打印成型纳米羟基磷灰石/壳聚糖/聚己内酯三元复合支架材料的构建及表征

文题释义： 组织工程骨：将体外培养的功能相关的种子细胞种植于天然的或人工合成的支架材料内,加入生长因子体外培养一段时间,将他们移植到体内,促进组织修复和骨再生的人工骨。组织工程骨形成的3要素为：支架材料、成骨细胞、生长因子。 生物陶瓷：生物表面活性陶瓷通常含有羟基,还可做成多孔性,生物组织可长入并同其表面发生牢固的键合;生物吸收性陶瓷的特点是能部分吸收或者全部吸收,在生物体内能诱发新生骨的生长。生物活性陶瓷具有骨传导性,它作为一个支架,成骨在其表面进行;还可作为多种物质的外壳或填充骨缺损。生物陶瓷有羟基磷灰石陶瓷、磷酸三钙陶瓷等。  背景：目前常用的骨缺损修复支架材料种类较多,但单一类型材料难以满足骨组织工程支架材料的要求,通过合适的方法将几种单一材料组合形成复合型材料,综合考虑各种材料优缺点,是近年来学者们的研究重点。 目的：构建纳米羟基磷灰石/壳聚糖/聚己内酯三元复合支架材料,并作表征分析研究。 方法：采用3D打印成型技术制备纳米羟基磷灰石/壳聚糖/聚己内酯多孔三元复合支架材料,从X射线衍射分析、吸水率、抗压强度、体外降解性能、孔径分析、扫描电镜分析等多个维度对支架材料进行表征研究。 结果与结论：①X射线衍射分析显示,纳米羟基磷灰石/壳聚糖/聚己内酯多孔三元复合支架的晶型峰图与羟基磷灰石粉末衍射标准卡片类似,表明该三元复合支架是通过物理作用相互结合的,不影响羟基磷灰石的生物学功能;②三元复合支架的吸水率为18.28%,亲水性好,支架可承受的最大压力为1 415 N,其体外降解速率与成骨速率相当;③显微镜下可见三元复合支架的内孔为方形,孔径250 µm,孔径大小均匀、分布有致;④扫描电镜下三元复合支架可见,壳聚糖和聚己内酯组成的纤维排列整齐有序,成网格状, 羟基磷灰石呈颗粒状在纤维表面均匀分布,三元复合材料呈现均匀、疏松的微孔结构;⑤结果表明,通过3D打印成型技术可成功制备纳米羟基磷灰石/壳聚糖/聚己内酯三元复合支架材料,其具有适度的抗压强度、一定的孔隙率、适宜的降解速度和吸水率,能为修复骨缺损的奠定基础。 ORCID: 0000-0002-6321-9160(余和东) 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程       

Influence of processing parameters on pore structure of 3D porous chitosan-alginate polyelectrolyte complex scaffolds

Fabrication of porous polymeric scaffolds with controlled structure can be challenging. In this study, we investigated the influence of key experimental parameters on the structures and mechanical properties of resultant porous chitosan-alginate (CA) polyelectrolyte complex (PEC) scaffolds, and on proliferation of MG-63 osteoblast-like cells, targeted at bone tissue engineering. We demonstrated that the porous structure is largely affected by the solution viscosity, which can be regulated by the acetic acid and alginate concentrations. We found that the CA PEC solutions with viscosity below 300 Pa.s yielded scaffolds of uniform pore structure and that more neutral pH promoted more complete complexation of chitosan and alginate, yielding stiffer scaffolds. CA PEC scaffolds produced from solutions with viscosities below 300 Pa.s also showed enhanced cell proliferation compared with other samples. By controlling the key experimental parameters identified in this study, CA PEC scaffolds of different structures can be made to suit various tissue engineering applications.     

Image analysis of the axonal ingrowth into poly(D,L-lactide) porous scaffolds in relation to the 3-D porous structure

Porous polymer scaffolds are promising materials for neural tissue engineering because they offer valuable three-dimensional (3-D) supports for the in vitro and in vivo axonal growth and tissue expansion. At the time being, how the in vivo neuronal cell development depends on the scaffold 3-D architecture is unknown. Therefore, scanning electron micrographs of longitudinal sections of porous polylactide scaffolds and immunohistological sections of these scaffolds after implantation and neurofilament staining have been studied by image analysis. Pore orientation and axonal ingrowth have been investigated by spectral analysis on gray level SEM images. Binary image processing has been carried out and the binary images have been studied by spectral analysis in order to estimate the possible effect of the image noise on the real pattern. In addition to axonal orientation, density and length distribution of the regenerated axons into the polymer scaffold have been measured. Dependence of the axonal ingrowth on the 3D-polymer scaffold has been discussed on the basis of the collected data.     

Quantitative biorelevant profiling of material microstructure within 3D porous scaffolds via multiphoton fluorescence microscopy

This study presents a novel approach, based on fluorescence multiphoton microscopy (MPM), to image and quantitatively characterize the microstructure and cell-substrate interactions within microporous scaffold substrates fabricated from synthetic biodegradable polymers. Using fluorescently dyed scaffolds fabricated from poly(DTE carbonate)/poly(DTO carbonate) blends of varying porosity and complementary green fluorescent protein-engineered fibroblasts, we reconstructed the three-dimensional distribution of the microporous and macroporous regions in 3D scaffolds, as well as cellular morphological patterns. The porosity, pore size and distribution, strut size, pore interconnectivity, and orientation of both macroscale and microscale pores of 3D scaffolds were effectively quantified and validated using complementary imaging techniques. Compared to other scaffold characterizing techniques such as confocal imaging and scanning electron microscopy (SEM), MPM enables the acquisition of images from scaffold thicknesses greater than a hundred microns with high signal-to-noise ratio, reduced bulk photobleaching, and the elimination of the need for deconvolution. In our study, the morphology and cytoskeletal organization of cells within the scaffold interior could be tracked with high resolution within the limits of penetration of MPM. Thus, MPM affords a promising integrated platform for imaging cell-material interactions within the interior of polymeric biomaterials.