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Carboxymethyl Hyaluronan-Stabilized Nanoparticles for Anticancer Drug Delivery.

Woodman JL, Suh MS, Zhang J, Kondaveeti Y, Burgess DJ, White BA, Prestwich GD, Kuhn LT - Int J Cell Biol (2015)

Bottom Line: CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days.Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP.This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Connecticut, Storrs, CT 06269, USA ; Department of Reconstructive Sciences, UConn Health, Farmington, CT 06030, USA.

ABSTRACT
Carboxymethyl hyaluronic acid (CMHA) is a semisynthetic derivative of HA that is recognized by HA binding proteins but contains an additional carboxylic acid on some of the 6-hydroxyl groups of the N-acetyl glucosamine sugar units. These studies tested the ability of CMHA to stabilize the formation of calcium phosphate nanoparticles and evaluated their potential to target therapy resistant, CD44(+)/CD24(-/low) human breast cancer cells (BT-474EMT). CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days. Surface plasmon resonance showed that nCaP(CMHA)CDDP binds to CD44, but less effectively than CMHA or hyaluronan. nCaP(CMHA-AF488) was taken up by CD44(+)/CD24(-) BT-474EMT breast cancer cells within 18 hours. nCaP(CMHA)CDDP was as cytotoxic as free CDDP against the BT-474EMT cells. Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP. This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

No MeSH data available.


Related in: MedlinePlus

Physical characterization of nCaPCMHA using X-ray diffraction and transmission electron microscopy. (a) XRD spectra of nCaPCMHA suspension (nCaPCMHA wet), lyophilized nCaPCMHA (nCaPCMHA dry), and lyophilized CaP without stabilizer added during precipitation (microCaP dry). Hydroxyapatite standard (JCPDS, #09-0432, bars) is shown for comparison. nCaPCMHA both wet and dry has a major broad peak corresponding to the major peaks of hydroxyapatite. MicroCaP pattern has major peaks characteristic of brushite (peaks denoted by open circles). MicroCaP was precipitated without a stabilizer and has larger crystalline particles (narrow peaks). With the CMHA stabilizer present the crystallization is halted, depicted by broad peaks with little long range order. (b) TEM image of nCaPCMHACDDP showing particles 20–50 nm in diameter clustered into larger, 200 nm, particles due to drying prior to imaging.
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fig2: Physical characterization of nCaPCMHA using X-ray diffraction and transmission electron microscopy. (a) XRD spectra of nCaPCMHA suspension (nCaPCMHA wet), lyophilized nCaPCMHA (nCaPCMHA dry), and lyophilized CaP without stabilizer added during precipitation (microCaP dry). Hydroxyapatite standard (JCPDS, #09-0432, bars) is shown for comparison. nCaPCMHA both wet and dry has a major broad peak corresponding to the major peaks of hydroxyapatite. MicroCaP pattern has major peaks characteristic of brushite (peaks denoted by open circles). MicroCaP was precipitated without a stabilizer and has larger crystalline particles (narrow peaks). With the CMHA stabilizer present the crystallization is halted, depicted by broad peaks with little long range order. (b) TEM image of nCaPCMHACDDP showing particles 20–50 nm in diameter clustered into larger, 200 nm, particles due to drying prior to imaging.

Mentions: The physical characteristics of nCaPCMHACDDP are shown in Table 1. Precipitation of nCaPCMHA resulted in an efficient yield of 2.3 ± 0.4 mg per mL of precipitation solution. The CDDP concentration in the nCaPCMHACDDP suspension was 4.1 ± 1.4 mg/mL, with a drug loading of 140 ± 12 μg CDDP/mg nCaPCMHA. nCaPCMHACDDP were on average 204 ± 13 nm in diameter as measured by dynamic light scattering with a polydispersity of 0.116. The zeta potential of the particles was −43 ± 4 mV, which is greater than the colloidal stability threshold of ±30 mV thereby providing evidence of enhanced stability due to surface charge prevention of aggregation [31]. XRD data showed nCaPCMHA was poorly crystalline apatite based on the major peak occurring at 30° which corresponds well with hydroxyapatite. MicroCaP made without the CMHA stabilizer was more crystalline in nature resembling brushite and poorly crystalline hydroxyapatite (Figure 2(a)) [32, 33]. The introduction of CMHA clearly restricts the crystallization of CaP, as can be seen by the broad peaks of nCaPCMHA XRD spectra relative to the spectra of microCaP which exhibited a crystalline pattern with major peaks of hydroxyapatite as well as brushite (calcium hydrogen phosphate dehydrate, CaHPO4·2H2O). Transmission electron microscopy (TEM) (Figure 2(b)) revealed that nCaPCMHACDDP in suspension forms small aggregates that correlate well with their measured particle size using DLS, 204 ± 13 nm. The addition of CMHA during precipitation of CaP resulted in successful stabilization of nCaP. TEM images revealed small 30–80 nm particles, agglomerated into larger particles, which likely accounts for the 200 nm size measured by DLS. These characteristics of the nCaPCMHACDDP resulted in a clog-free injectable nanoparticle suspension via a 25G needle.


Carboxymethyl Hyaluronan-Stabilized Nanoparticles for Anticancer Drug Delivery.

Woodman JL, Suh MS, Zhang J, Kondaveeti Y, Burgess DJ, White BA, Prestwich GD, Kuhn LT - Int J Cell Biol (2015)

Physical characterization of nCaPCMHA using X-ray diffraction and transmission electron microscopy. (a) XRD spectra of nCaPCMHA suspension (nCaPCMHA wet), lyophilized nCaPCMHA (nCaPCMHA dry), and lyophilized CaP without stabilizer added during precipitation (microCaP dry). Hydroxyapatite standard (JCPDS, #09-0432, bars) is shown for comparison. nCaPCMHA both wet and dry has a major broad peak corresponding to the major peaks of hydroxyapatite. MicroCaP pattern has major peaks characteristic of brushite (peaks denoted by open circles). MicroCaP was precipitated without a stabilizer and has larger crystalline particles (narrow peaks). With the CMHA stabilizer present the crystallization is halted, depicted by broad peaks with little long range order. (b) TEM image of nCaPCMHACDDP showing particles 20–50 nm in diameter clustered into larger, 200 nm, particles due to drying prior to imaging.
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Related In: Results  -  Collection

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fig2: Physical characterization of nCaPCMHA using X-ray diffraction and transmission electron microscopy. (a) XRD spectra of nCaPCMHA suspension (nCaPCMHA wet), lyophilized nCaPCMHA (nCaPCMHA dry), and lyophilized CaP without stabilizer added during precipitation (microCaP dry). Hydroxyapatite standard (JCPDS, #09-0432, bars) is shown for comparison. nCaPCMHA both wet and dry has a major broad peak corresponding to the major peaks of hydroxyapatite. MicroCaP pattern has major peaks characteristic of brushite (peaks denoted by open circles). MicroCaP was precipitated without a stabilizer and has larger crystalline particles (narrow peaks). With the CMHA stabilizer present the crystallization is halted, depicted by broad peaks with little long range order. (b) TEM image of nCaPCMHACDDP showing particles 20–50 nm in diameter clustered into larger, 200 nm, particles due to drying prior to imaging.
Mentions: The physical characteristics of nCaPCMHACDDP are shown in Table 1. Precipitation of nCaPCMHA resulted in an efficient yield of 2.3 ± 0.4 mg per mL of precipitation solution. The CDDP concentration in the nCaPCMHACDDP suspension was 4.1 ± 1.4 mg/mL, with a drug loading of 140 ± 12 μg CDDP/mg nCaPCMHA. nCaPCMHACDDP were on average 204 ± 13 nm in diameter as measured by dynamic light scattering with a polydispersity of 0.116. The zeta potential of the particles was −43 ± 4 mV, which is greater than the colloidal stability threshold of ±30 mV thereby providing evidence of enhanced stability due to surface charge prevention of aggregation [31]. XRD data showed nCaPCMHA was poorly crystalline apatite based on the major peak occurring at 30° which corresponds well with hydroxyapatite. MicroCaP made without the CMHA stabilizer was more crystalline in nature resembling brushite and poorly crystalline hydroxyapatite (Figure 2(a)) [32, 33]. The introduction of CMHA clearly restricts the crystallization of CaP, as can be seen by the broad peaks of nCaPCMHA XRD spectra relative to the spectra of microCaP which exhibited a crystalline pattern with major peaks of hydroxyapatite as well as brushite (calcium hydrogen phosphate dehydrate, CaHPO4·2H2O). Transmission electron microscopy (TEM) (Figure 2(b)) revealed that nCaPCMHACDDP in suspension forms small aggregates that correlate well with their measured particle size using DLS, 204 ± 13 nm. The addition of CMHA during precipitation of CaP resulted in successful stabilization of nCaP. TEM images revealed small 30–80 nm particles, agglomerated into larger particles, which likely accounts for the 200 nm size measured by DLS. These characteristics of the nCaPCMHACDDP resulted in a clog-free injectable nanoparticle suspension via a 25G needle.

Bottom Line: CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days.Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP.This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Connecticut, Storrs, CT 06269, USA ; Department of Reconstructive Sciences, UConn Health, Farmington, CT 06030, USA.

ABSTRACT
Carboxymethyl hyaluronic acid (CMHA) is a semisynthetic derivative of HA that is recognized by HA binding proteins but contains an additional carboxylic acid on some of the 6-hydroxyl groups of the N-acetyl glucosamine sugar units. These studies tested the ability of CMHA to stabilize the formation of calcium phosphate nanoparticles and evaluated their potential to target therapy resistant, CD44(+)/CD24(-/low) human breast cancer cells (BT-474EMT). CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days. Surface plasmon resonance showed that nCaP(CMHA)CDDP binds to CD44, but less effectively than CMHA or hyaluronan. nCaP(CMHA-AF488) was taken up by CD44(+)/CD24(-) BT-474EMT breast cancer cells within 18 hours. nCaP(CMHA)CDDP was as cytotoxic as free CDDP against the BT-474EMT cells. Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP. This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

No MeSH data available.


Related in: MedlinePlus