Silica Biomineralization with Lignin Involves Si-O-C Bonds That Stabilize Radicals.

Plants undergo substantial biomineralization of silicon, which is deposited primarily in cell walls as amorphous silica. The mineral formation could be moderated by the structure and chemistry of lignin, a polyphenol polymer that is a major constituent of the secondary cell wall. However, the reactions between lignin and silica have not yet been well elucidated. Here, we investigate silica deposition onto a lignin model compound. Polyphenyl propanoid was synthesized from coniferyl alcohol by oxidative coupling with peroxidase in the presence of acidic tetramethyl orthosilicate, a silicic acid precursor. Raman, Fourier transform infrared, and X-ray photoelectron spectroscopies detected changes in lignin formation in the presence of silicic acid. Bonds between the Si-O/Si-OH residues and phenoxyl radicals and lignin functional groups formed during the first 3 h of the reaction, while silica continued to form over 3 days. Thermal gravimetric analysis indicated that lignin yields increased in the presence of silicic acid, possibly via the stabilization of phenolic radicals. This, in turn, resulted in shorter stretches of the lignin polymer. Silica deposition initiated within a lignin matrix via the formation of covalent Si-O-C bonds. The silica nucleants grew into 2-5 nm particles, as observed via scanning transmission electron microscopy and energy-dispersive X-ray spectroscopy. Additional silica precipitated into an extended gel. Collectively, our results demonstrate a reciprocal relation by which lignin polymerization catalyzes the formation of silica, and at the same time silicic acid enhances lignin polymerization and yield.

Siliplant1 B-domain precipitates silica spheres, aggregates, or gel, depending on Si-precursor to peptide ratios.

Silica is extensively deposited by plants, however, only little is known about the molecular control over this process. Siliplant1 is the only known plant protein to precipitate biosilica. The protein contains seven repeats made of three domains. One of the domains exhibits a conserved sequence, which catalyzes silica precipitation in vitro. Here, silica was synthesized by the activity of a peptide carrying this conserved sequence. Infrared spectroscopy and thermal gravimetric analyses showed that the peptide was bound to the mineral. Scanning electron microscopy showed that silica-peptide particles of 22 ± 4 nm aggregated to spherical structures of 200-300 nm when the ratio of silicic acid to the peptide was below 183:1 molecules. When the ratio was about 183:1, similar particles aggregated into irregular structures, and silica gel formed at higher ratios. Solid-state NMR spectra indicated that the irregular aggregates were richer in Si-O-Si bonds as well as disordered peptide. Our results suggest that the peptide catalyzed the condensation of silicic acid and the formation of ∼20 nm particles, which aggregated into spheres. Excess of the peptide stabilized surface Si-OH groups that prevented spontaneous Si-O-Si bonding between aggregates. Under Si concentrations relevant to plant sap, the peptide and possibly Siliplant1, could catalyze nucleation of silica particles that aggregate into spherical aggregates.

Biocompatibility and bioactivity of hardystonite-based nanocomposite scaffold for tissue engineering applications.

Bone injury, especially bone damages due to the removal of bone tumors, is one of the most important issues in the field of therapeutic research in tissue engineering applications. In this context, ceramic-based composites have attracted widespread attention since they have mechanical properties close to the natural bone, hence providing similar conditions for the extracellular matrix (ECM). Thus, in this study, hardystonite and diopside (HT-Di) scaffolds containing various diopside amounts from 5 to 25 wt% were prepared by the space holder method. The results revealed that the fabricated scaffolds contain 70%-75% porosity with a pore size of 300-500 µm and a compressive strength of about 0.54 to 1.71 MPa which is perfectly in the range of the compressive strength of the sponge bone. Noticeably, great apatite formation ability was observed in the scaffold with diopside, although the scaffold without diopside showed poor bioactivity. The MTT assay depicted that the inclusion of diopside into hardystonite scaffold resulted in dramatic enhancement in the MG-63 cell viability. Moreover, the scaffold with diopside offered greater cell attachment and spreading than the scaffold without diopside. Therefore, the synergistic effects of the scaffold with 12.5 wt% of diopside, including great mechanical characteristic, excellent bioactivity, and appealing biocompatibility enable it to be an appealing choice for bone tissue engineering applications.

Co-incorporation of strontium and fluorine into diopside scaffolds: Bioactivity, biodegradation and cytocompatibility evaluations.

This study focuses on the effect of Sr-, F-, and their co-doping on the structure, biodegradation, bioactivity and cytocompatibility of diopside-based scaffolds, using X-ray diffraction, Raman spectroscopy, field-emission scanning electron microscopy, Archimedes densitometry, inductively coupled plasma spectroscopy, pH-metry, and cell MTT assay. The structural characterization of the scaffolds confirmed the successful incorporation of the dopants into the ceramic. In addition, all the doped scaffolds presented higher apatite-forming ability levels in comparison to the undoped one, where the highest and the least impact of doping on bioactivity belonged to F- and co-doping, respectively. It was found that the biodegradation difference of the scaffolds in terms of principal ions and the chance of F-incorporation into precipitated apatite determine the bioactivity difference of the samples. Osteoblast-like MG-63 cells exhibited the highest and lowest compatibility to the Sr-doped and co-doped scaffolds, respectively. In summary, F- and Sr-doping offered the highest bioactivity and cytocompatibility, respectively, whereas co-doping presented the weakest behaviors comparatively.

A comparative study on biological properties of novel nanostructured monticellite-based composites with hydroxyapatite bioceramic.

In this research, novel monticellite/hydroxyapatite (HA) ceramic composites were successfully prepared by mechanical method. The ability of nanostructured monticellite-based ceramic composites to form a suitable bond to living hard tissues, and stimulate osteoblast-like cells proliferation may be different for various ratios of the reinforcement to monticellite matrix. The differences in physico-chemical characteristics, bone-like apatite formation, cytocompatibility, cell viability and in vitro osteogenic activity of nanostructured monticellite/HA ceramic composites were explored. The surface reactivity and bioactivity of the composite samples were evaluated in vitro in simulated body fluid (SBF). A comparative time- and dose-dependent MTT test showed that the ions release from nanostructured monticellite/HA composites dissolution significantly stimulated cell proliferation and growth than control at a certain concentration range. The cells viability exposed to the composite extract was higher than control and mineral material of bone (HA), illustrating that cytocompatibility was improved due to the presence of magnesium (Mg) and silicon (Si) elements in the composite structure. The comparative results of alkaline phosphatase (ALP) bioactivity assay showed that the osteogenic proliferation of osteoblast-like G292 cell was increased more by the ceramic composites extract than control. These comparative results demonstrated that nanostructured monticellite-based ceramic composites possessed good in vitro bioactivity, cytocompatibility and osteogenic properties, and may be utilized as the promising bioactive materials for bone tissue regeneration and replacement.

Silicic Acid-Mediated Formation of Tannic Acid Nanocoatings.

Tannic acid (TA) adheres to a broad variety of different materials and forms versatile surface coatings for technical and biological applications. In mild alkaline conditions, autoxidation processes occur and a firm monolayer is formed. Up to now, thicker coatings are obtained in only a cross-linked multilayer fashion. This study presents an alternative method to form continuous TA coatings using orthosilicic acid (Siaq). Adsorption kinetics and physical properties of TA coatings in the presence of Siaq were determined using a quartz-crystal microbalance and nanoplasmonic spectroscopy. An in situ TA layer thickness of 200 nm was obtained after 24 h in solutions supplemented with 80 µM Siaq. Dry-state measurements indicated a highly hydrated layer in situ. Furthermore, chemical analysis by Fourier transform infrared spectroscopy revealed possible complexation of TA by Siaq, whereas UV-vis spectroscopy did not indicate an interaction of Siaq in the autoxidation process of TA. Investigation of additional metalloid ions showed that germanic acid was also able to initiate a continuous coating formation of TA, whereas boric acid prevented the polymerization process. In comparison to that of TA, the coating formation of pyrogallol (PG) and gallic acid (GA) was not affected by Siaq. PG formed continuous coatings also without Siaq, whereas GA formed only a monolayer in the presence of Siaq. However, Siaq induced a continuous layer formation of ellagic acid. These results indicate the specific importance of orthosilicic acid in the coating formation of polyphenolic molecules with multiple ortho-dihydroxy groups and open new possibilities to deposit TA on interfaces.

Silicification process in diatom algae using different silicon chemical sources: Colloidal silicic acid interactions at cell surface.

The silicon transport and use inside cells are key processes for understanding how diatoms metabolize this element in the silica biogenic cycle in the ocean. A spin-probe electron paramagnetic resonance (EPR) study over time helped to investigate the interacting properties and the internalization mechanisms of silicic acid from different silicon sources into the cells. Diatom cells were grown in media containing biogenic amorphous substrates, such as diatomaceous earth and sponge spicules, and crystalline sodium metasilicate. It was found that the amorphous biogenic silicon slowed down the internalization process probably due to formation of colloidal particles at the cell surface after silicic acid condensation. Weaker interactions occurred with sponge spicules silicon source if compared to the other sources. The EPR results were explained by analyzing transcript level changes of silicon transporters (SITs) and silaffins (SILs) in synchronized Thalassiosira pseudonana cultures over time. The results indicated that the transport role of SITs is minor for silicic acid from both biogenic and crystalline substrates, and the role of SIT3 is linked to the transport of silicon inside the cells, mainly in the presence of sponge spicules. SIL3 transcripts were expressed in the presence of all silicon sources, while SIL1 transcripts only with sponge spicules. The data suggest that the transport of silicic acid from various silicon sources in diatoms is based on different physico-chemical interactions with the cell surface.

Secondary structure and dynamics study of the intrinsically disordered silica-mineralizing peptide P5 S3 during silicic acid condensation and silica decondensation.

The silica forming repeat R5 of sil1 from Cylindrotheca fusiformis was the blueprint for the design of P5 S3 , a 50-residue peptide which can be produced in large amounts by recombinant bacterial expression. It contains 5 protein kinase A target sites and is highly cationic due to 10 lysine and 10 arginine residues. In the presence of supersaturated orthosilicic acid P5 S3 enhances silica-formation whereas it retards the dissolution of amorphous silica (SiO2 ) at globally undersaturated concentrations. The secondary structure of P5 S3 during these 2 processes was studied by circular dichroism (CD) spectroscopy, complemented by nuclear magnetic resonance (NMR) spectroscopy of the peptide in the absence of silicate. The NMR studies of dual-labeled (13 C, 15 N) P5 S3 revealed a disordered structure at pH 2.8 and 4.5. Within the pH range of 4.5-9.5 in the absence of silicic acid, the CD data showed a disordered structure with the suggestion of some polyproline II character. Upon silicic acid polymerization and during dissolution of preformed silica, the CD spectrum of P5 S3 indicated partial transition into an α-helical conformation which was transient during silica-dissolution. The secondary structural changes observed for P5 S3 correlate with the presence of oligomeric/polymeric silicic acid, presumably due to P5 S3 -silica interactions. These P5 S3 -silica interactions appear, at least in part, ionic in nature since negatively charged dodecylsulfate caused similar perturbations to the P5 S3 CD spectrum as observed with silica, while uncharged ß-d-dodecyl maltoside did not affect the CD spectrum of P5 S3 . Thus, with an associated increase in α-helical character, P5 S3 influences both the condensation of silicic acid into silica and its decondensation back to silicic acid.

Silica nanoparticles as sources of silicic acid favoring wound healing in vitro.

There is good evidence that certain silicon-containing materials promote would healing and their common feature is the delivery of orthosilicic acid (Si(OH)4) either directly or following metabolism. In this respect, amorphous silica nanoparticles (NP), which dissolve in aqueous environments releasing up to 2mM orthosilicic acid, may be appropriate 'slow release' vehicles for bioactive silicon. Here we studied the impact of silica NP suspensions (primary particles∼10nm) in undersaturated conditions (below 2mM Si) with differing degrees of surface charge and dissolution rate on human dermal fibroblasts (CCD-25SK cells) viability, proliferation and migration in a cellular wound model. Silica was shown to be non-toxic for all forms and concentrations tested and whilst the anticipated stimulatory effect of orthosilicic acid was observed, the silica NPs also stimulated fibroblast proliferation and migration. In particular, the amine-functionalized particles promoted wound closure more rapidly than soluble orthosilicic acid alone. We suggest that this effect is related to easy cellular internalization of these particles followed by their intracellular dissolution releasing silicic acid at a faster rate than its direct uptake from the medium. Our findings indicate that amorphous silica-based NPs may favour the delivery and release of bioactive silicic acid to cells, promoting wound healing.

Intrafibrillar silicified collagen scaffold modulates monocyte to promote cell homing, angiogenesis and bone regeneration.

The immunomodulatory functions of monocytes are increasingly being recognized. Silicified collagen scaffolds (SCSs), produced by infiltrating collagen matrices with intrafibrillar amorphous silica, exhibit osteogenic and angiogenic potential and are promising candidates in tissue engineering. Here, we demonstrate that SCS promotes in situ bone regeneration and angiogenesis via monocyte immunomodulation. Increased numbers of TRAP-positive monocytes, nestin-positive bone marrow stromal cells (BMSCs) and CD31-positive and endomucin-positive new vessels can be identified from new bone formation regions in a murine calvarial defect model. In addition, sustained release of silicic acid by SCS stimulates differentiation of blood-derived monocytes into TRAP-positive cells, with increased expressions of SDF-1α, TGF-ß1, VEGFa and PDGF-BB. These cytokines further promote homing of BMSCs and endothelial progenitor cells as well as neovascularization. Taken together, these novel findings indicate that SCSs possess the ability to enhance recruitment of progenitor cells and promote osteogenesis and angiogenesis by immunomodulation of monocytes.

Biocompatibility and bioactivity of porous polymer-derived Ca-Mg silicate ceramics.

Magnesium is a trace element in the human body, known to have important effects on cell differentiation and the mineralisation of calcified tissues. This study aimed to synthesise highly porous Ca-Mg silicate foamed scaffolds from preceramic polymers, with analysis of their biological response. Akermanite (Ak) and wollastonite-diopside (WD) ceramic foams were obtained from the pyrolysis of a liquid silicone mixed with reactive fillers. The porous structure was obtained by controlled water release from selected fillers (magnesium hydroxide and borax) at 350°C. The homogeneous distribution of open pores, with interconnects of modal diameters of 160-180µm was obtained and maintained after firing at 1100°C. Foams, with porosity exceeding 80%, exhibited compressive strength values of 1-2MPa. In vitro studies were conducted by immersion in SBF for 21days, showing suitable dissolution rates, pH and ionic concentrations. Cytotoxicity analysis performed in accordance with ISO10993-5 and ISO10993-12 standards confirmed excellent biocompatibility of both Ak and WD foams. In addition, MC3T3-E1 cells cultured on the Mg-containing scaffolds demonstrated enhanced osteogenic differentiation and the expression of osteogenic markers including Collagen Type I, Osteopontin and Osteocalcin, in comparison to Mg-free counterparts. The results suggest that the addition of magnesium can further enhance the bioactivity and the potential for bone regeneration applications of Ca-silicate materials. STATEMENTS OF SIGNIFICANCE: Here, we show that the incorporation of Mg in Ca-silicates plays a significant role in the enhancement of the osteogenic differentiation and matrix formation of MC3T3-E1 cells, cultured on polymer-derived highly porous scaffolds. Reduced degradation rates and improved mechanical properties are also observed, compared to Mg-free counterparts, suggesting the great potential of Ca-Mg silicates as bone tissue engineering materials. Excellent biocompatibility of the new materials, in accordance to the ISO10993-5 and ISO10993-12 standard guidelines, confirms the preceramic polymer route as an efficient synthesis methodology for bone scaffolds. The use of hydrated fillers as porogens is an additional novelty feature presented in the manuscript.

In-vitro bioactivity, biocompatibility and dissolution studies of diopside prepared from biowaste by using sol-gel combustion method.

Diopside was synthesized from biowaste (Eggshell) by sol-gel combustion method at low calcination temperature and the influence of two different fuels (urea, l-alanine) on the phase formation temperature, physical and biological properties of the resultant diopside was studied. The synthesized materials were characterized by heating microscopy, FTIR, XRD, BET, SEM and EDAX techniques. BET analysis reveals particles were of submicron size with porosity in the nanometer range. Bone-like apatite deposition ability of diopside scaffolds was examined under static and circulation mode of SBF (Simulated Body Fluid). It was noticed that diopside has the capability to deposit HAP (hydroxyapatite) within the early stages of immersion. ICP-OES analysis indicates release of Ca, Mg, Si ions and removal of P ions from the SBF, but in different quantities from diopside scaffolds. Cytocompatability studies on human bone marrow stromal cells (hBMSCs) revealed good cellular attachment on the surface of diopside scaffolds and formation of extracellular matrix (ECM). This study suggests that the usage of eggshell biowaste as calcium source provides an effective substitute for synthetic starting materials to fabricate bioproducts for biomedical applications.

In vivo biocompatibility of custom-fabricated apatite-wollastonite-mesenchymal stromal cell constructs.

We have used the additive manufacturing technology of selective laser sintering (SLS), together with post SLS heat treatment, to produce porous three dimensional scaffolds from the glass-ceramic apatite-wollastonite (A-W). The A-W scaffolds were custom-designed to incorporate a cylindrical central channel to increase cell penetration and medium flow to the center of the scaffolds under dynamic culture conditions during in vitro testing and subsequent in vivo implantation. The scaffolds were seeded with human bone marrow mesenchymal stromal cells (MSCs) and cultured in spinner flasks. Using confocal and scanning electron microscopy, we demonstrated that MSCs formed and maintained a confluent layer of viable cells on all surfaces of the A-W scaffolds during dynamic culture. MSC-seeded, with and without osteogenic pre-differentiation, and unseeded A-W scaffolds were implanted subcutaneously in MF1 nude mice where osteoid formation and tissue in-growth were observed following histological assessment. The results demonstrate that the in vivo biocompatibility and osteo-supportive capacity of A-W scaffolds can be enhanced by SLS-custom design, without the requirement for osteogenic pre-induction, to advance their potential as patient-specific bone replacement materials.

Understanding the composition-structure-bioactivity relationships in diopside (CaO·MgO·2SiO2)-tricalcium phosphate (3CaO·P2O5) glass system.

The present work is an amalgamation of computation and experimental approach to gain an insight into composition-structure-bioactivity relationships of alkali-free bioactive glasses in the CaO-MgO-SiO2-P2O5 system. The glasses have been designed in the diopside (CaO·MgO·2SiO2; Di)-tricalcium phosphate (3CaO·P2O5; TCP) binary join by varying the Di/TCP ratio. The melt-quenched glasses have been investigated for their structure by molecular dynamic (MD) simulations as well as by nuclear magnetic resonance spectroscopy (NMR). In all the investigated glasses silicate and phosphate components are dominated by Q(2) (Si) and Q(0) (P) species, respectively. The apatite forming ability of the glasses was investigated using X-ray diffraction (XRD), infrared spectroscopy after immersion of glass powders in simulated body fluid (SBF) for time durations varying between 1 h and 14 days, while their chemical degradation has been studied in Tris-HCl in accordance with ISO 10993-14. All the investigated glasses showed good bioactivity without any substantial variation. A significant statistical increase in metabolic activity of human mesenchymal stem cells (hMSCs) when compared to the control was observed for Di-60 and Di-70 glass compositions under both basal and osteogenic conditions.

Na2SiF6:Cu,P: a new OSL phosphor for the radiation dosimetric applications.

A new Cu,P-doped, sodium fluorosilicate-based optically stimulated luminescence (OSL) phosphor is developed. This phosphor shows good OSL properties, and the sensitivity is comparable with that of the commercial Al(2)O(3):C (Landauer, Inc.) phosphor. For the luminescence averaged over initial 1 s, blue-stimulated luminescence and green-stimulated luminescence sensitivities were found to be 0.76 and 3.8 times, respectively, of Al(2)O(3):C (Landauer, Inc.) with 28 % of post-irradiation fading in 3 days and nil thereafter. The simple preparation procedure, fast decay, very good sensitivity and moderate fading will make this phosphor suitable for radiation dosimetry, using OSL.

Silicic acid competes for dimethylarsinic acid (DMA) immobilization by the iron hydroxide plaque mineral goethite.

A surface complexation modeling approach was used to extend the knowledge about processes that affect the availability of dimethylarsinic acid (DMA) in the soil rhizosphere in presence of a strong sorbent, e.g., Fe plaques on rice roots. Published spectroscopic and molecular modeling information suggest for the organoarsenical agent to form bidentate-binuclear inner-sphere surface complexes with Fe hydroxides similar to the inorganic As oxyanions. However, since also the ubiquitous silicic acid oxyanion form the same bidentate binuclear surface complexes, our hypothesis was that it may have an effect on the adsorption of DMA by Fe hydroxides in soil. Our experimental batch equilibrium data show that DMA is strongly adsorbed in the acidic pH range, with a steep adsorption edge in the circumneutral pH region between the DMA acidity constant (pKa=6.3) and the point of zero charge value of the goethite adsorbent (pHpzc=8.6). A 1-pK CD-MUSIC surface complexation model was chosen to fit the experimental adsorption vs. pH data. The same was done for silicic acid batch equilibrium data with our goethite adsorbent. Both model parameters for individual DMA and silicic acid adsorption were then merged into one CD-MUSIC model to predict the binary DMA+Si adsorption behavior. Silicic acid (500 µM) was thus predicted by the model to strongly compete for DMA with up to 60% mobilization of the latter at a pH6. This model result could be verified subsequently by experimental batch equilibrium data with zero adjustable parameters. The thus quantified antagonistic relation between DMA and silicic acid is discussed as one of factors to explain the increase of the DMA proportion in rice grains as observed upon silica fertilization of rice fields.

Una década de monitoreo de plomo en sangre en niños escolares del área metropolitana de Monterrey, NL / A decade of lead monitoring in schoolchildren in the metropolitan area of Monterrey NL, Mexico

Objetivo. Establecer las concentraciones de plomo (Pb) en sangre en niños escolares de 1998 y 2008, así como su asociación con factores de riesgo. Material y métodos. Se llevó a cabo un monitoreo de Pb en sangre de niños de entre 6 y 12 años que cursan educación primaria en 17 escuelas diferentes, ubicadas en distintas zonas del área metropolitana de Monterrey, de 1998 a 2008. Resultados. Se obtuvieron niveles séricos de 9.6 ± 3.0 (µg/dL rango de 3.18 a 20.88) en 1998 y de 4.5±4.8 µg/dL (rango de 3.3 a 53.7) en 2008, lo que mostró una disminución de 2.1 veces en nivel de Pb (p<0.01). Conclusiones. La reducción de los niveles séricos de Pb demuestran los mejores controles ambientales e industriales y probablemente el éxito de retirar el Pb de la gasolina durante los años noventa.

Objective. To establish the blood lead concentration and associated risk factors in schoolchildren during 1998 and 2008. Materials and methods. A blood lead screening was conducted in schoolchildren of 6-12 years of age, enrolled in 17 elementary schools of the metropolitan area of Monterrey, México, during 1998 and 2008. Results. The mean blood lead level were 9.6 ± 3.0 (µg/dL range of 3.18 to 20.88) in 1998 and 4.5±4.8 µg/dL (range of 3.3 to 53.7) showing a 2.1-times reduction in blood lead levels (p<0.01). Conclusions. This reduction in blood lead levels demonstrate environmental and industrial control improvements and the benefits of fading out the leaded gasoline during the 1990's.

Mechanical reinforcement of diopside bone scaffolds with carbon nanotubes.

Carbon nanotubes are ideal candidates for the mechanical reinforcement of ceramic due to their excellent mechanical properties, high aspect ratio and nanometer scale diameter. In this study, the effects of multi-walled carbon nanotubes (MWCNTs) on the mechanical properties of diopside (Di) scaffolds fabricated by selective laser sintering were investigated. Results showed that compressive strength and fracture toughness improved significantly with increasing MWCNTs from 0.5 to 2 wt %, and then declined with increasing MWCNTs to 5 wt %. Compressive strength and fracture toughness were enhanced by 106% and 21%, respectively. The reinforcing mechanisms were identified as crack deflection, MWCNTs crack bridging and pull-out. Further, the scaffolds exhibited good apatite-formation ability and supported adhesion and proliferation of cells in vitro.

Nanoparticle/Polymer assembled microcapsules with pH sensing property.

The dual-labeled microcapsules via nanoparticle/polymer assembly based on polyamine-salt aggregates can be fabricated for the ratiometric intracellular pH sensing. After deposition of SiO2 nanoparticles on the poly(allylamine hydrochloride)/multivalent anionic salt aggregates followed by silicic acid treatment, the generated microcapsules are stable in a wide pH range (3.0 ∼ 8.0). pH sensitive dye and pH insensitive dye are simultaneously labeled on the capsules, which enable the ratiometric pH sensing. Due to the rough and positively charged surface, the microcapsules can be internalized by several kinds of cells naturally. Real-time measurement of intracellular pH in several living cells shows that the capsules are all located in acidic organelles after being taken up. Furthermore, the negatively charged DNA and dyes can be easily encapsulated into the capsules via charge interaction. The microcapsules with combination of localized pH sensing and drug loading abilities have many advantages, such as following the real-time transportation and processing of the carriers in cells.